1

ROLE OF CIRCULAR RNAs AS DIAGNOSTIC BIOMARKERS

IN EARLY DETECTION OF CANCER: A
REVIEW REPORT

SUBMITTED BY:
NIMRA SHAHEEN

DEPARTMENT OF ZOOLOGY
LAHORE COLLEGE FOR WOMEN UNIVERSITY,
LAHORE 2026
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ROLE OF CIRCULAR RNAs AS DIAGNOSTIC BIOMARKERS

IN
EARLY DETECTION OF CANCER: A REVIEW REPORT

A REPORT SUBMITED TO LAHORE COLLEGE FOR WOMEN

UNIVERSITY IN PARTIAL FULFILMENT OF REQUIREMENT
FOR THE AWARD OF DEGREE IN BS ZOOLOGY
BY
NIMRA SHAHEEN
2225138038
2022-2026

DEPARTMENT OF ZOOLOGY

LAHORE COLLEGE FOR WOMEN UNIVERSITY, LAHORE

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2026
CERTIFICATE
This is to certify that the work described in this literature review report submitted by Ms.
Nimra Shaheen to Department of Zoology, Lahore College for Women University has
been carried out under my supervision. I also certify that the enclosed manuscript has
been prepared under my supervision and I endorse its evaluation for the award of a BS
degree through the official procedure of the University.

Pro. Dr. Farzana Rashid

LCWU, Lahore
Date:
Verified By:

Pro. Dr. Farzana Rashid

Chairperson
Department of Zoology
Stamp

Controller of Examination
Stamp
 4

Date:
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1. INTRODUCTION

1.1 Discovery of circular RNAs

Circular RNAs (circRNAs) are a class of endogenous RNA molecules characterized by a
covalently closed loop structure without 5′ caps and 3′ polyadenylated tails. circular
RNA molecules were first observed years ago in plant viroids and later in eukaryotic
cells, they were initially considered rare splicing errors or nonfunctional byproducts of
aberrant RNA processing. The limited sensitivity of early sequencing technologies
contributed to the underestimation of their abundance and biological relevance. The
advent of high-throughput RNA sequencing (RNA-seq) and improved bioinformatics
algorithms around 2012 led to the rediscovery of circRNAs as widespread and conserved
molecules in eukaryotic transcriptomes. Comprehensive transcriptome analyses revealed
that circRNAs are abundantly expressed in human and animal tissues, often in a tissue-
specific and developmental stage-specific manner, indicating regulated biogenesis rather
than random formation (Salzman et al., 2013). Subsequent studies demonstrated that
circRNAs are evolutionarily conserved across species and can accumulate to significant
levels in certain cell types, particularly in neuronal tissues. Their stable accumulation and
distinct expression patterns strongly suggest functional importance rather than
transcriptional noise (Jeck et al., 2014).

1.2 Structural Characteristics of Circular RNAs

The defining structural feature of circRNAs is their covalently closed continuous loop,
which is generated through a noncanonical splicing event known as back-splicing. In this
process, a downstream 5′ splice donor site joins to an upstream 3′ splice acceptor site,
forming a circular molecule instead of a linear transcript. This structure distinguishes
circRNAs from traditional linear messenger RNAs (mRNAs) (Chen et al., 2015).
Because circRNAs lack free ends, they are resistant to exonuclease-mediated
degradation, such as digestion by RNase R. This resistance results in significantly longer
half-lives compared to linear RNAs, making circRNAs highly stable molecules within
cells and extracellular fluids. Their enhanced stability is one of the most important
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characteristics that underlies their potential use as clinical biomarkers. Furthermore,

circRNAs often display cell type-specific, tissue-specific, and disease-specific expression
profiles. Many circRNAs are expressed at levels comparable to or even higher than their
corresponding linear transcripts, indicating that circularization is a regulated and
biologically meaningful process (Salzman et al., 2016).

1.3 Biogenesis of Circular RNAs

CircRNAs are generated from precursor mRNA (pre-mRNA) transcripts through a back-
splicing mechanism that competes with canonical splicing. Several molecular
mechanisms contribute to circRNA formation, including intron pairing-driven
circularization. Complementary sequences within flanking introns, such as Al repeats in
humans, facilitate the bringing together of splice sites to promote circularization (Zhang
et al., 2014). Another mechanism involves RNA-binding proteins (RBPs), which bind to
flanking intronic regions and act as bridges to facilitate circular RNA formation. Proteins
such as Quaking (QKI) and Muscleblind (MBL) have been shown to enhance circRNA
biogenesis, demonstrating that circRNA production is actively regulated at the post-
transcriptional level (Conn et al., 2015). In addition, exon skipping can produce lariat
intermediates containing exons that are further processed into circular RNAs. This lariat-
driven circularization mechanism highlights the complexity of circRNA formation and its
close relationship with alternative splicing pathways (Ashwal-Fluss et al.,2014).

1.4 Classification of Circular RNAs

Based on their composition and origin, circRNAs are categorized into three primary
types. The most abundant are exonic circRNAs (EcircRNAs), which consist exclusively
of exon sequences and are predominantly located in the cytoplasm. These circRNAs are
mainly involved in post-transcriptional gene regulation (Memczak et al., 2013). Circular
intronic RNAs (ciRNAs) are derived solely from intronic sequences and are typically
retained within the nucleus. They have been implicated in the regulation of transcription
of their parental genes, suggesting a role in nuclear gene expression control (Zhang et al.,
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2013). Exon–intron circRNAs (EIciRNAs) contain both exonic and intronic sequences
and are also primarily localized in the nucleus. EIciRNAs have been shown to interact
with RNA polymerase II and enhance transcriptional activity of their host genes,
indicating functional diversity among circRNA subclasses (Li et al.,2015).

1.5 Biological Functions of Circular RNAs

One of the most extensively studied functions of circRNAs is their role as microRNA
(miRNA) sponges. CircRNAs can contain multiple binding sites for specific miRNAs,
allowing them to sequester these molecules and prevent them from repressing their target
mRNAs. A classical example is ciRS-7 (CDR1as), which contains numerous binding
sites for miR-7 and modulates its regulatory network (Hansen et al., 2013). In addition to
miRNA sponging, circRNAs can interact directly with RNA-binding proteins,
functioning as scaffolds or decoys that modulate protein activity and cellular signaling
pathways. These interactions further expand their regulatory capacity within the cell
(Abdelmohsen et al.,2017).
Although circRNAs are generally classified as non-coding RNAs, recent evidence
indicates that some circRNAs can undergo cap-independent translation through internal
ribosome entry sites (IRES) or N6-methyladenosine (m6A) modifications. This discovery
broadens the functional spectrum of circRNAs beyond traditional gene regulation
(Pamudurti et al., 2017). CircRNAs participate in diverse biological processes that
influence cancer development and progression. One of their most extensively studied
functions is acting as microRNA (miRNA) sponges. By binding to complementary
miRNA sequences, circRNAs can sequester miRNAs and prevent them from inhibiting
their target messenger RNAs. This circRNA–miRNA–mRNA regulatory axis plays a
critical role in controlling gene expression programs associated with tumor growth,
apoptosis resistance, angiogenesis, and metastasis. (Hansen et al., 2013). Beyond miRNA
sponging, circRNAs can interact with RNA-binding proteins (RBPs), serving as scaffolds
that modulate protein localization and function. Some circRNAs have been shown to
regulate transcription of their parental genes or compete with canonical splicing events.
Recent evidence also indicates that certain circRNAs contain internal ribosome entry
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sites (IRES) and open reading frames, enabling them to encode functional peptides.
These diverse regulatory mechanisms demonstrate that circRNAs are active participants
in oncogenic signaling rather than passive molecular by-products. (Kristensen et al.,
2019).

Figure 1:Circular RNAs characteristics, biogenesis, mechanisms and functions in liver

cancer

1.7 Why circRNAs Are Ideal Diagnostic Biomarkers

Circular RNAs possess several biological features that make them superior biomarker
candidates compared with linear RNAs. Their covalently closed loop structure protects
them from exonuclease-mediated degradation, giving them exceptional stability in body
fluids such as blood, saliva, urine, and cerebrospinal fluid. Because of this stability,
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circRNAs remain intact even under harsh storage or processing conditions, allowing
accurate detection using clinical assays. (Li et al., 2022). In addition, circRNAs exhibit
tissue-specific and developmental stage-specific expression patterns. This specificity
enables researchers to identify particular circRNAs that correlate with distinct cancer
types or tumor stages. Studies have demonstrated that abnormal circRNA expression is
associated with carcinogenesis, tumor progression, metastasis, and drug resistance,
indicating their potential value not only in diagnosis but also in prognosis and therapeutic
monitoring. (Li et al., 2025). Another important property is their presence in extracellular
vesicles, especially exosomes. Exosomal circRNAs are released into circulation and can
be detected non-invasively through liquid biopsy approaches. Reviews analyzing multiple
studies have reported that exosomal circRNAs can detect cancers with high sensitivity
and specificity, highlighting their clinical utility as minimally invasive diagnostic
biomarkers. (Zhang et al., 2023).

1.8 circRNAs in Blood-Based Cancer Diagnosis

Blood-based biomarkers are particularly valuable because they allow repeated sampling
without invasive procedures. Systematic reviews analyzing dozens of studies on
colorectal cancer have shown that circRNAs detected in blood or tissue samples exhibit
strong diagnostic potential. In one comprehensive review of 55 studies, circRNAs
demonstrated promising performance as non-invasive markers capable of distinguishing
cancer patients from healthy individuals. (Latifi-Pakdehi et al., 2024). The diagnostic
utility of circulating circRNAs arises from their release into the bloodstream via tumor
cell apoptosis, necrosis, or active secretion. Because tumor cells display dysregulated
circRNA expression profiles, these molecules serve as molecular fingerprints of
malignancy. Detection methods such as quantitative reverse transcription PCR (qRT-
PCR), RNA sequencing, and microarray analysis allow precise quantification of circRNA
levels, facilitating their clinical application. (Latifi-Pakdehi et al., 2024).
Moreover, circRNAs can distinguish between cancer stages and subtypes. Differential
expression analysis shows that certain circRNAs are significantly upregulated in early
tumors compared with normal tissue, indicating their value in early detection. These
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findings support the use of circRNA panels rather than single markers to enhance
diagnostic accuracy. (Zhang et al., 2023; Li et al., 2025).

1.9 Cancer-Specific Diagnostic Applications

CircRNAs have been investigated across a wide spectrum of cancers, including breast,
colorectal, lung, gastric, liver, and hematological malignancies. In breast cancer,
dysregulated circRNA expression has been linked to tumor initiation, progression,
metastasis, and resistance to therapy. These molecular changes provide a basis for using
circRNAs as biomarkers for diagnosis, classification of molecular subtypes, and
prediction of treatment responses. (Li et al., 2025). Similarly, in colorectal cancer,
circRNAs identified through systematic review analyses were consistently associated
with disease development and progression. Because these molecules can be measured in
blood samples, they are particularly promising for screening populations at risk.
Researchers emphasize that circRNAs may serve as complementary markers alongside
existing screening tools to improve diagnostic sensitivity. (Latifi-Pakdehi et al., 2024)
CircRNAs also play functional roles in tumor biology, further supporting their biomarker
potential. For example, some circRNAs act as oncogenes that promote malignant
phenotypes, while others function as tumor suppressors that inhibit proliferation or
inflammation. Such dual regulatory roles reflect their involvement in key cancer
pathways and highlight their diagnostic relevance. (Li et al., 2022).

1.10 Clinical Advantages Over Traditional Biomarkers

Compared with protein-based markers, circRNAs offer several clinical advantages. First,
they show higher stability and resistance to degradation. Second, they can be detected at
low concentrations using highly sensitive molecular assays. Third, circRNA signatures
often reflect tumor-specific molecular alterations rather than general physiological
changes, reducing false-positive results. These advantages collectively make circRNAs
attractive candidates for early cancer screening programs. (Zhang et al., 2023). Another
benefit is their compatibility with liquid biopsy technologies. Liquid biopsy enables
cancer detection through analysis of circulating tumor DNA, RNA, or cells in body
fluids. CircRNAs are particularly suitable for this approach because they are abundant in
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extracellular vesicles and remain stable during circulation. As a result, circRNA-based

assays may enable real-time monitoring of tumor progression and treatment response. (Li
et al., 2022; Zhang et al., 2023).

1.11 Global Burden of Cancer

Cancer continues to represent one of the most significant global health challenges,
accounting for millions of new diagnoses and deaths annually. Despite advancements in
surgical techniques, chemotherapy, radiotherapy, targeted therapy, and immunotherapy,
overall survival rates remain highly dependent on the stage at which cancer is diagnosed.
Early-stage tumors are often curable, whereas advanced metastatic disease carries a poor
prognosis. However, many cancers remain asymptomatic during early stages, leading to
delayed diagnosis and limited therapeutic success. This reality highlights the urgent need
for highly sensitive and specific molecular biomarkers capable of detecting malignancies
at their earliest stages, ideally through non-invasive or minimally invasive methods.
(Bray et al., 2018).
Traditional diagnostic biomarkers such as carcinoembryonic antigen (CEA), alpha-
fetoprotein (AFP), prostate-specific antigen (PSA), and CA19-9 suffer from limited
sensitivity and specificity, particularly in early-stage disease. Imaging modalities, while
useful, may fail to detect microscopic or pre-invasive lesions. Tissue biopsies remain the
gold standard for diagnosis but are invasive, costly, and not always feasible for repeated
monito13ring. Consequently, research has increasingly focused on molecular biomarkers
derived from nucleic acids, including circulating DNA and various classes of noncoding
RNAs. Among these, circular RNAs (circRNAs) have emerged as promising candidates
due to their distinctive structural and functional properties. (Siravegna et al., 2017).

1.12 Unveiling Circular RNAs

Circular RNAs were initially observed several decades ago but were long considered
splicing errors or transcriptional artifacts. The development of high-throughput RNA
sequencing and advanced bioinformatics algorithms has dramatically reshaped this view,
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revealing that circRNAs are abundant, conserved, and dynamically regulated molecules
across eukaryotic species. They are generated through a noncanonical splicing process
known as back-splicing, in which a downstream splice donor joins to an upstream splice
acceptor, forming a covalently closed circular transcript. This process can involve
exoncircularization, intron circularization, or exon-intron combinations. (Chen et al.,
2020)
Unlike linear RNAs, circRNAs lack free 5′ and 3′ ends, which protects them from
exonuclease-mediated degradation. This structural feature significantly enhances their
half-life within cells and extracellular fluids. Studies have demonstrated that circRNAs
are more stable than their linear mRNA counterparts under conditions of RNase treatment
or prolonged storage. Such remarkable stability makes circRNAs particularly suitable for
biomarker development, especially in liquid biopsy applications where RNA degradation
is a common challenge. (Jeck and Sharpless, 2014).

1.13 Stability and Detectability in Body Fluids

One of the most compelling attributes of circRNAs as diagnostic biomarkers is their

detectability in various body fluids, including plasma, serum, saliva, urine, and
exosomes. Their circular configuration renders them resistant to RNase degradation,
allowing reliable quantification even in extracellular environments. CircRNAs have been
detected in circulating exosomes—small extracellular vesicles that protect RNA
molecules during transport between cells—further enhancing their diagnostic potential in
non-invasive testing. (Li et al., 2015). The presence of circRNAs in exosomes is
particularly significant for early cancer detection because tumor cells actively release
exosomes containing tumor-specific molecular signatures into circulation. Analysis of
exosomal circRNA profiles can therefore reflect underlying tumor biology without
requiring tissue biopsy. This capability supports the development of liquid biopsy
platforms aimed at detecting cancer at preclinical stages. (Memczak et al., 2013).

1.14 CircRNAs in Specific Cancer Types

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In gastric cancer, circRNAs have shown significant diagnostic promise. Multiple studies
have identified dysregulated circRNAs in both tumor tissues and plasma samples of
patients. Diagnostic performance analyses have demonstrated that certain circRNAs
achieve higher area under the curve (AUC) values compared to traditional markers such
as CEA and CA19-9. Importantly, some circRNAs exhibit altered expression even in
early-stage gastric cancer, highlighting their utility in screening strategies (Chen et al.,
2017).
In hepatocellular carcinoma (HCC), circRNAs such as circMTO1 and circZKSCAN1
have been reported to be significantly dysregulated. Their expression levels correlate with
tumor size, vascular invasion, and clinical stage. Plasma circRNA levels have shown
diagnostic value in distinguishing HCC patients from individuals with chronic liver
disease, suggesting potential use in high-risk populations such as patients with hepatitis B
or C infection. (Qin et al., 2016). Colorectal cancer (CRC) research has similarly
revealed numerous circRNAs with diagnostic potential. Several circRNAs demonstrate
stage-specific expression patterns, and combined circRNA panels have achieved strong
sensitivity and specificity in distinguishing early CRC from healthy controls. These
findings are particularly important given the rising global incidence of CRC in younger
populations. (Hsiao et al., 2017).

1.14 Evidence from Meta-Analyses

Comprehensive meta-analyses integrating data from multiple independent studies have

confirmed the diagnostic value of circRNAs across diverse malignancies. Pooled
analyses typically report moderate to high sensitivity and specificity, with AUC values
often exceeding 0.80. Such findings indicate strong discriminatory ability between cancer
patients and non-cancer individuals. Importantly, subgroup analyses frequently
demonstrate superior diagnostic accuracy when circRNAs are detected in plasma rather
than tissue samples. (Vo et al., 2019). Meta-analytic evidence also suggests that
combining multiple circRNAs into biomarker panels enhances diagnostic performance
compared to single-marker approaches. Multiplex circRNA signatures leverage the
heterogeneity of cancer biology and reduce false-positive or false-negative rates. This
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panel-based strategy aligns with precision medicine approaches that emphasize

comprehensive molecular profiling for accurate disease detection. (Wang et al., 2018).

Figure 2: The Landscape of Circular RNA in Cancer Cell

1.15 CircRNAs in Early-Stage Cancer Detection

The ultimate goal of biomarker research is detection of cancer at its earliest, most
treatable stage. CircRNAs are particularly promising in this context because their
dysregulation often occurs during early tumorigenesis. Studies have demonstrated altered
circRNA expression in stage I cancers, including lung, gastric, and colorectal cancers.
Early-stage diagnostic AUC values comparable to or exceeding those of traditional
biomarkers suggest that circRNAs may serve as early warning indicators of malignant
transformation. (Shang et al., 2019). Furthermore, longitudinal monitoring studies
indicate that circRNA levels may change dynamically in response to tumor progression
or therapeutic intervention. Such temporal responsiveness supports their use not only in
early detection but also in monitoring disease recurrence or minimal residual disease after
treatment. (Zhang et al., 2018).
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1.16 Advantages Over Other Biomarkers

Compared with circulating tumor DNA (ctDNA), circRNAs offer enhanced stability and
may be present at higher abundance in early disease stages. While ctDNA detection
depends on tumor DNA shedding, which may be minimal in early tumors, circRNAs can
reflect active transcriptional dysregulation even before substantial DNA release occurs.
Additionally, circRNA detection methods such as quantitative reverse transcription PCR
(qRT-PCR) and next-generation sequencing are increasingly standardized and cost-
effective. (Wan et al., 2017). Unlike protein biomarkers that may be influenced by
inflammation or non-malignant conditions, circRNAs often display tumor-specific
expression patterns. Their cell-type specificity increases diagnostic precision and reduces
confounding effects. Such advantages position circRNAs as complementary or
potentially superior molecular tools for cancer screening. (Li et al., 2018).

1.17 Future Perspectives

Future research directions include integration of circRNA profiles with other omics
datasets such as DNA methylation, proteomics, and metabolomics. Machine learning
algorithms may further enhance predictive modeling based on circRNA expression
signatures. With continued technological refinement and large multicenter validation
trials, circRNAs hold strong potential to revolutionize early cancer detection and improve
global cancer outcomes. (Chen et al., 2020). Advances in sequencing technologies,
bioinformatics, and machine learning are expected to accelerate circRNA biomarker
discovery. High-throughput RNA sequencing combined with computational analysis
enables identification of novel circRNAs and evaluation of their diagnostic performance
across large datasets. Integrating circRNA profiles with other molecular markers may
further improve early detection accuracy. (Latifi-Pakdehi et al., 2024).
Future clinical applications may involve circRNA-based diagnostic panels that detect
cancer before symptoms appear. Such tools could transform cancer screening by enabling
early intervention, reducing mortality, and improving patient outcomes. Continued
research focusing on validation, standardization, and mechanistic understanding will be
essential to realize this potential. (Zhang et al., 2023; Li et al., 2025). Circular RNAs
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(circRNAs) are increasingly recognized as promising molecular biomarkers because their

covalently closed structure provides exceptional resistance to exonuclease degradation,
allowing them to remain stable in blood, saliva, urine, and extracellular vesicles. This
stability makes circRNAs highly suitable for non-invasive diagnostic testing and
longitudinal disease monitoring. Furthermore, their evolutionary conservation and tissue-
specific expression patterns suggest that circRNAs are biologically functional molecules.
(Sun et al., 2021).
Abnormal circRNA expression has been documented across multiple cancer types, where
they participate in tumor initiation, progression, invasion, and metastasis through
modulation of gene regulatory networks. Importantly, many circRNAs exhibit cancer-
type specificity, enabling them to distinguish malignant tissues from normal tissues with
greater precision than conventional biomarkers. Such specificity indicates that circRNAs
could serve as molecular signatures that reflect tumor origin and pathological state.
(Zhang et al., 2021).Quantitative meta-analyses evaluating diagnostic performance
demonstrate that circRNAs possess measurable clinical accuracy as cancer biomarkers.
One pooled analysis reported an overall area under the ROC curve (AUC) of
approximately 0.79, along with moderate sensitivity and specificity values. These results
suggest that circRNA detection assays may function effectively as complementary
diagnostic tools alongside imaging a tests (Wang et al., 2018).
Further statistical synthesis indicates that panels containing multiple circRNAs
outperform single-marker assays. Combined circRNA signatures have achieved
sensitivity values approaching 0.89, specificity near 0.94, and AUC values around 0.97,
indicating excellent discriminative power between cancer patients and healthy individuals
(Li et al., 2019). Cancer-specific analyses provide additional evidence supporting
circRNA-based diagnostics. In gastric cancer studies, multiple circRNAs were
consistently dysregulated, and pooled statistical evaluation produced a diagnostic odds
ratio of 8.78 with an AUC of 0.81. Notably, circRNAs detected in plasma samples
showed higher diagnostic accuracy than those measured in tissue biopsies, highlighting
their potential for minimally invasive screening strategies (Jiang et al., 2019).
Certain individual circRNAs also demonstrate strong clinical relevance. For example,
circSMARCA5 is frequently downregulated in diverse malignancies such as
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hepatocellular carcinoma, lung cancer, glioblastoma, and colorectal cancer. Reduced

expression of this circRNA correlates with aggressive tumor characteristics and poor
patient survival, indicating its dual value as both a diagnostic and prognostic biomarker
(Zhang et al., 2021). High-throughput sequencing studies further support circRNA utility
in early detection. Multi-omics analysis in ovarian cancer identified numerous disease-
associated circRNAs and constructed regulatory networks linking circRNAs,
microRNAs, and mRNAs. A selected panel combining circRNA and miRNA markers
demonstrated perfect discrimination between early-stage cancer patients and healthy
individuals within the analyzed dataset, illustrating the potential of integrated molecular
diagnostic models.
(Liu et al., 2025).

1. 18 Limitations and Challenges

Despite promising results, the clinical translation of circRNA biomarkers faces several
challenges. Many studies conducted so far involve small sample sizes and lack validation
in large multicenter clinical trials. Without large-scale validation, it is difficult to
establish standardized diagnostic thresholds or confirm reproducibility across
populations. (Li et al., 2025). Another limitation is methodological variability.
Differences in sample preparation, RNA extraction techniques, and detection platforms
can lead to inconsistent results across studies. Standardized protocols are therefore
necessary to ensure reliability and comparability of circRNA measurements before they
can be widely adopted in clinical diagnostics. (Latifi-Pakdehi et al., 2024).
Finally, the biological mechanisms underlying many circRNAs remain poorly
understood. Although numerous circRNAs have been identified as potential biomarkers,
only a subset has been functionally characterized. Further research is required to clarify
their molecular roles and to determine which candidates have the highest clinical value.
(Li et al., 2025)
Despite encouraging evidence, several challenges must be addressed before circRNAs
can be routinely implemented in clinical diagnostics. These include the need for
standardized extraction protocols, normalization strategies, large-scale validation cohorts,
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and reproducible detection platforms. Inter-study variability in sample handling and data
analysis currently limits comparability across studies. (Kristensen et al., 2019).

Table: Role of Circular RNAs as Diagnostic Biomarkers in Early Detection of Cancer

Relevance to Early Cancer
Aspect Description
Detection

CircRNAs possess a covalently

closed-loop structure lacking 5′ High stability in body fluids
Structural
and 3′ ends, making them enables reliable detection even
Characteristics
resistant to exonuclease in early-stage disease.
degradation.

Formed through back-splicing of

Cancer-related alterations in
pre-mRNA, involving exon
Biogenesis splicing mechanisms lead to
circularization, intron
Mechanism tumor-specific circRNA
circularization, or exon–intron
expression patterns.
combinations.

More stable than linear RNAs Allows accurate measurement

(mRNA, miRNA) due to in plasma, serum, saliva,
Molecular Stability
resistance to RNase R urine, and exosomes for non-
degradation. invasive screening.

Enhances diagnostic
Exhibit tissue-specific and
Expression specificity and reduces false-
disease-specific expression
Specificity positive results compared to
profiles.
conventional protein markers.

Supports minimally invasive

Presence in Liquid Detectable in circulating blood, early detection and repeated
Biopsy exosomes, and other body fluids. monitoring of high-risk
individuals.

Act as miRNA sponges, interact Reflect early oncogenic

Functional with RNA-binding proteins, signaling changes, making
Mechanisms regulate transcription, and in them sensitive indicators of
some cases encode peptides. malignant transformation.

Evaluated using sensitivity, Many circRNAs show AUC >

Diagnostic
specificity, diagnostic odds ratio 0.80 in early-stage cancers,
Performance
(DOR), and area under ROC indicating strong
Metrics
curve (AUC). discriminative ability.
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Gastric cancer, colorectal cancer,

Demonstrated diagnostic value
hepatocellular carcinoma, lung
Cancer Types across multiple malignancies,
cancer, breast cancer, pancreatic
Studied increasing clinical
cancer, oral squamous cell
applicability.
carcinoma.

Comparison with More stable than ctDNA and Potentially superior sensitivity
Traditional protein markers such as CEA, in early-stage disease
Biomarkers CA19-9, AFP. detection.

Multiple circRNAs combined into Multiplex circRNA signatures

Combination
panels improve diagnostic increase sensitivity and
Biomarker Panels
accuracy. specificity for early detection.

Non-invasive sampling, high Suitable for population

Clinical
stability, reproducibility, dynamic screening and early-stage
Advantages
monitoring capability. tumor identification.

Lack of standardized detection Requires large-scale validation

Challenges protocols, small sample sizes in before routine clinical
studies, inter-study variability. implementation.

Integration with multi-omics data May enhance precision

Future Directions and AI-based predictive screening and personalized
modeling. cancer diagnostics.
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AIMS AND OBJECTIVES

 To assess the potential of circular RNAs (circRNAs) as effective diagnostic
biomarkers in the early detection of various cancers.
 To examine the biological properties of circRNAs, including their stability,
abundance, and tissue-specific expression patterns.
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LITERATURE REVIEW

Wang et al., 2016 examined that circular RNAs (circRNAs) have emerged as a
significant class of endogenous noncoding RNAs characterized by their covalently
closed-loop structure, which distinguishes them from linear RNAs. This circular
configuration confers remarkable resistance to exonuclease-mediated degradation,
resulting in enhanced stability and longer half-life in comparison to linear transcripts.
Advances in high-throughput RNA sequencing technologies have revealed that circRNAs
are not rare splicing by-products but rather abundant, conserved, and dynamically
regulated molecules across diverse species. Importantly, circRNAs often demonstrate
tissue-specific and developmental-stage-specific expression patterns, suggesting tightly
controlled regulatory functions. These features collectively make circRNAs highly
attractive candidates for molecular research and potential clinical applications.
Functionally, circRNAs participate in multiple regulatory mechanisms that influence
gene expression and cellular homeostasis. One of the most extensively studied roles of
circRNAs is their ability to act as microRNA (miRNA) sponges. By binding and
sequestering specific miRNAs, circRNAs prevent miRNAs from suppressing their target
messenger RNAs, thereby indirectly modulating protein expression. In addition to
miRNA sponging, circRNAs can interact with RNA-binding proteins (RBPs), serving as
protein scaffolds or decoys that influence transcription, RNA stability, and translation.
Emerging evidence also suggests that certain circRNAs may regulate parental gene
transcription or even encode functional peptides. These diverse biological roles highlight
the importance of circRNAs in maintaining cellular regulatory networks and underscore
their potential impact in pathological [Link] studies have increasingly linked
circRNA dysregulation to the initiation and progression of various diseases, particularly
cancer. Aberrant expression of specific circRNAs has been observed in numerous
malignancies, where they contribute to tumorigenesis by regulating oncogenes, tumor
suppressor genes, and signaling pathways involved in proliferation, apoptosis, invasion,
and metastasis. Notably, circRNAs have been detected in body fluids such as blood,
saliva, and exosomes, demonstrating their stability and accessibility in non-invasive
clinical samples. This presence in readily obtainable biological fluids enhances their
potential as diagnostic and prognostic biomarkers. Collectively, current literature
supports the growing recognition of circRNAs as promising molecular tools for early
cancer detection, prognosis assessment, and the development of targeted therapeutic
strategies.

Lü et al., 2017 described that Recent research has increasingly focused on the role of
endogenous noncoding circular RNAs (circRNAs) in the molecular pathogenesis of
breast cancer. Although circRNAs have been widely implicated in various malignancies,
their comprehensive expression patterns in breast cancer were not fully characterized
until genome-wide profiling studies were conducted. Using high-throughput circRNA
 22

microarray technology, investigators were able to construct a detailed circRNA

expression landscape, revealing significant dysregulation in tumor tissues compared to
normal controls. In the referenced two-stage study, 1,155 circRNAs were identified as
differentially expressed, with 715 upregulated and 440 downregulated in breast cancer
tissues. This large-scale alteration in circRNA expression suggests that circRNAs are
actively involved in tumor biology rather than representing incidental transcriptional by-
[Link] validate the microarray findings, selected candidate circRNAs were examined
using quantitative real-time polymerase chain reaction (qRT-PCR), confirming consistent
expression patterns. Specifically, hsa_circ_103110, hsa_circ_104689, and
hsa_circ_104821 were significantly upregulated, whereas hsa_circ_006054,
hsa_circ_100219, and hsa_circ_406697 were markedly downregulated in breast cancer
tissues. Bioinformatic analyses further demonstrated that these circRNAs contain
complementary miRNA response elements, indicating their potential function as
competing endogenous RNAs (ceRNAs) capable of sequestering tumor-associated
microRNAs. Through these interactions, circRNAs may regulate key oncogenic signaling
pathways that drive cellular proliferation, invasion, and metastasis in breast
[Link], the diagnostic value of circRNAs was also evaluated in this study.
When three downregulated circRNAs (hsa_circ_006054, hsa_circ_100219, and
hsa_circ_406697) were combined as a biomarker panel, the area under the receiver
operating characteristic (ROC) curve reached 0.82, demonstrating good discriminatory
power between breast cancer and non-cancer tissues. This finding underscores the clinical
relevance of circRNAs as potential diagnostic biomarkers. Collectively, the study
provides compelling evidence that circRNAs are significantly dysregulated in breast
cancer and contribute to carcinogenesis through miRNA sequestration and pathway
modulation, highlighting their promise for early detection and molecular-targeted
strategies.

Yang et al., 2020 revealed that Circular RNAs (circRNAs) have emerged as a significant
class of highly conserved and stable non-coding RNAs characterized by their covalently
closed-loop structure. This structural configuration provides resistance to exonuclease-
mediated degradation, resulting in enhanced stability compared to linear RNAs.
Functionally, circRNAs participate in diverse regulatory mechanisms, including
modulation of gene transcription and protein translation. They achieve these roles
primarily through interactions with RNA-binding proteins and by acting as microRNA
(miRNA) sponges, thereby influencing post-transcriptional gene regulation. These
multifaceted biological functions suggest that circRNAs are integral components of
cellular regulatory networks and may play important roles in maintaining normal cellular
[Link] the context of gastric cancer, accumulating evidence indicates that
circRNAs are aberrantly expressed and contribute to tumorigenesis. Dysregulated
circRNAs have been shown to influence critical cancer-related processes such as cell
proliferation, apoptosis, migration, invasion, and metastasis. Through their ability to
regulate oncogenes and tumor suppressor genes—often via circRNA–miRNA–mRNA
interaction networks—circRNAs can modulate key signaling pathways involved in
gastric cancer progression. This regulatory influence highlights their potential
mechanistic role in carcinogenesis and supports the concept that circRNA expression
profiles reflect underlying molecular alterations within tumor [Link] their
 23

biological significance, circRNAs hold considerable promise for clinical application in

gastric cancer. Their inherent stability and detectability in tissues and body fluids make
them attractive candidates for diagnostic biomarkers. Moreover, because circRNAs
participate in tumor-promoting pathways, they may also serve as therapeutic targets,
either by modulating their expression or disrupting their interaction networks.
Collectively, current literature underscores the dual diagnostic and therapeutic potential
of circRNAs in gastric cancer, emphasizing their relevance in future translational
research and precision oncology strategies.

Ojha et al., 2018 reported that Circular RNAs (circRNAs) are evolutionarily conserved
noncoding RNA molecules that naturally occur within the eukaryotic transcriptome.
Generated through a back-splicing mechanism, circRNAs form covalently closed-loop
structures that lack free 5′ and 3′ ends, rendering them resistant to exonuclease-mediated
degradation. This structural stability allows circRNAs to persist in various biological
environments, including serum, plasma, and tissue samples, making them highly suitable
for molecular profiling studies. Beyond structural uniqueness, circRNAs possess the
ability to interact with nucleic acids and ribonucleoprotein complexes, positioning them
as important regulators of gene expression. Their interactions contribute to the
modulation of transcriptional and post-transcriptional processes, thereby integrating
circRNAs into complex cellular regulatory [Link] research has demonstrated
that aberrant expression of circRNAs is associated with a wide spectrum of human
diseases, including neurological disorders, infectious diseases, and cancers. Differential
circRNA expression can be identified through advanced profiling techniques such as
RNA sequencing, microarrays, and quantitative real-time PCR, enabling detection in both
tissue and circulating body fluids. For example, specific circRNAs have been shown to
exhibit disease-specific expression patterns: hsa_circRNA_001937 is upregulated in
pulmonary tuberculosis, whereas hsa_circRNA_102101 is downregulated. In oncology,
hsa_circ_000178 has been reported to be upregulated in breast cancer and associated with
disease progression. These findings indicate that circRNA dysregulation reflects
underlying pathological processes and may serve as disease-specific molecular
[Link] notable is the abundance of circRNAs in mammalian brain tissue,
where they appear to play critical roles in neuronal development and function. Altered
circRNA expression has been linked to neurological conditions such as epilepsy, in
which Circ-EFCAB2 is significantly upregulated. The stability, tissue specificity, and
detectability of circRNAs in accessible biological fluids collectively support their
potential as noninvasive biomarkers for early disease detection and monitoring. Overall,
current literature highlights circRNAs as promising diagnostic tools across diverse
pathological conditions, underscoring their expanding relevance in translational medicine
and biomarker research.

Xia et al., 2018 Circular RNAs (circRNAs) represent a recently recognized class of
noncoding RNAs distinguished by their covalently closed-loop structure, which
differentiates them from conventional linear RNAs. This circular configuration renders
them resistant to exonuclease-mediated degradation, particularly by RNase R, thereby
 24

conferring remarkable stability within cells and body fluids. CircRNAs are abundantly
expressed in eukaryotic transcriptomes and exhibit high evolutionary conservation as
well as developmental stage–specific expression patterns. These unique biological
features suggest that circRNAs are not merely by-products of splicing events but
functionally relevant molecules with regulatory significance. Their stability and
persistence in biological samples provide a strong rationale for investigating their clinical
[Link] research over recent years has established a strong association
between circRNA dysregulation and various human diseases, particularly cancer.
Numerous studies have reported aberrant circRNA expression profiles across different
tumor types, indicating their involvement in oncogenesis and tumor progression.
Mechanistically, circRNAs contribute to cancer development by regulating key genes and
signaling pathways that control cell proliferation, apoptosis, migration, and invasion.
Through interactions with microRNAs, RNA-binding proteins, and transcriptional
machinery, circRNAs participate in complex regulatory networks that influence tumor
behavior. The consistent observation of cancer-specific circRNA expression patterns
further strengthens their relevance in malignancy [Link], circRNAs are not
confined to intracellular compartments; they can be released from tissues and cells into
extracellular environments, including blood and saliva. Their abundance and stability in
these body fluids highlight their potential as noninvasive biomarkers for cancer diagnosis
and prognosis. Because early detection is critical for improving cancer outcomes, the
identification of reliable molecular markers is of significant clinical importance. The
growing body of literature supports the concept that circRNAs may serve as promising
diagnostic and prognostic indicators, paving the way for future translational research
aimed at integrating circRNA-based assays into clinical oncology practice.
Cui et al., 2018 examined that Circular RNAs (circRNAs) are a novel class of
endogenous RNAs distinguished by their covalently closed-loop structure, which lacks
both 5′–3′ polarity and a polyadenylated tail. This structural uniqueness contributes to
their exceptional stability and resistance to exonuclease-mediated degradation. Advances
in high-throughput sequencing technologies have revealed that circRNAs are not rare
transcriptional artifacts but are abundantly expressed, evolutionarily conserved, and often
exhibit tissue-specific and developmental stage–specific expression patterns. These
 25

characteristics suggest that circRNAs have important physiological roles and are tightly
regulated within cellular [Link], circRNAs participate in diverse
biological processes that are closely associated with tumorigenesis and metastasis. One of
their most widely recognized functions is acting as competing endogenous RNAs
(ceRNAs) or microRNA (miRNA) sponges, thereby modulating post-transcriptional gene
regulation. In addition to miRNA sequestration, circRNAs can interact with RNA-
binding proteins, influencing transcription, RNA stability, and protein translation.
Emerging evidence also indicates that certain circRNAs possess protein-coding potential,
expanding their functional repertoire beyond noncoding regulatory roles. Through these
mechanisms, circRNAs contribute to critical cellular processes such as proliferation,
apoptosis, invasion, and metastatic progression in various [Link] their stability,
specificity, and detectability in tissues and body fluids, circRNAs have gained attention
as promising biomarkers for cancer diagnosis. Differential expression patterns of
circRNAs have been identified across multiple tumor types, underscoring their potential
clinical relevance. By reflecting underlying molecular alterations within cancer cells,
circRNAs may serve as sensitive indicators for early detection, disease monitoring, and
prognosis assessment. Overall, current literature highlights the expanding understanding
of circRNA classification, biogenesis, mechanisms, and functional implications in cancer,
supporting their potential integration into future diagnostic and translational oncology
strategies.

Cui et al., 2022 proclaimed that Circular RNAs (circRNAs) are a distinct class of single-
stranded endogenous RNAs generated through a noncanonical back-splicing process of
precursor mRNAs. Unlike linear RNAs, circRNAs form covalently closed-loop structures
without 5′ caps or 3′ polyadenylated tails. Their biogenesis primarily occurs through three
mechanisms: intron-pairing-driven circularization, RNA-binding protein (RBP)-mediated
circularization, and lariat-driven circularization. These mechanisms enable the precise
and regulated production of circRNAs within eukaryotic cells. While the majority of
circRNAs are localized in the cytoplasm, certain intron-containing circRNAs are retained
in the nucleus, where they may participate in transcriptional regulation. The complexity
of their formation and localization patterns underscores their functional diversity within
cellular [Link], circRNAs have been recognized as important regulators of
 26

gene expression. One of their most well-established roles is acting as microRNA

(miRNA) sponges, where they sequester miRNAs and prevent them from suppressing
target messenger RNAs. Additionally, circRNAs interact with RNA-binding proteins,
influencing protein function, RNA stability, and gene transcription. Emerging evidence
also suggests that some circRNAs possess protein-coding potential, further expanding
their biological significance. Through these mechanisms, circRNAs contribute to critical
processes involved in cancer development and progression, including cell proliferation,
apoptosis, invasion, and metastasis. Their tissue-specific and developmental stage–
specific expression patterns further highlight their regulatory importance in both normal
physiology and disease [Link] remarkable stability and longer half-life of circRNAs
compared to linear RNAs make them particularly attractive for clinical
[Link] are readily detectable in tissues and in various body fluids such as
plasma, saliva, and urine, supporting their feasibility for use in minimally invasive liquid
biopsy approaches. Accumulating studies have demonstrated that aberrant circRNA
expression correlates with cancer diagnosis, progression, and prognosis. Consequently,
circRNAs are increasingly recognized as promising biomarkers with significant potential
for early cancer detection and personalized medicine. Overall, current research
emphasizes the structural characteristics, functional roles, and translational relevance of
circRNAs, reinforcing their emerging value in oncology and clinical diagnostics.

Yang et al., 2017 Circular RNAs (circRNAs) are a recently characterized class of
endogenous non-coding RNAs distinguished by their covalently closed-loop structure,
which provides remarkable structural stability and resistance to exonuclease degradation.
They are highly abundant and evolutionarily conserved across mammalian species,
suggesting functional importance rather than being mere by-products of splicing events.
CircRNAs exhibit tissue-specific and cell-type-specific expression patterns, indicating
regulated biogenesis and specialized biological roles. Within eukaryotic cells, circRNAs
contribute to the regulation of gene expression at both transcriptional and post-
transcriptional levels, primarily through interactions with microRNAs (miRNAs), RNA-
binding proteins, and other regulatory [Link] evidence has linked circRNAs
to a wide range of human diseases, including atherosclerosis, neurological disorders,
diabetes, and particularly cancer. In oncological contexts, dysregulated circRNA
expression has been shown to influence tumor initiation and progression by modulating
key cellular processes such as proliferation, apoptosis, angiogenesis, invasion, and
metastasis. Mechanistically, circRNAs often function as competing endogenous RNAs
(ceRNAs), sequestering miRNAs and thereby regulating downstream target genes.
Additionally, circRNAs have been implicated in the modulation of major signal
transduction pathways, including those involved in cell survival and growth, further
highlighting their significance in disease [Link] a clinical perspective, the
stability, specificity, and detectability of circRNAs in tissues and body fluids position
them as promising diagnostic and prognostic biomarkers. Their aberrant expression
profiles in different cancers correlate with tumor stage, metastasis, and patient survival
outcomes. Consequently, circRNAs hold substantial potential for improving early cancer
detection and guiding therapeutic decision-making. Overall, recent advances in circRNA
research underscore their biological importance and clinical relevance, particularly in the
context of cancer diagnosis, prognosis, and targeted therapy development.
 27

Sheng et al., 2020 studied that Ovarian cancer (OC) remains one of the most aggressive
gynecologic malignancies, characterized by high invasiveness, metastatic potential, and
poor overall survival rates. Despite advances in cytoreductive surgery and platinum-
based chemotherapy, the prognosis for many patients remains unfavorable due to late-
stage diagnosis and frequent recurrence. The absence of highly sensitive and specific
biomarkers for early detection significantly contributes to delayed diagnosis and limited
therapeutic success. Consequently, there is an urgent need to identify novel molecular
markers that can improve early diagnosis, predict prognosis, and serve as potential
therapeutic targets in [Link] RNAs (circRNAs), a class of highly conserved, stable,
and abundant noncoding RNAs, have recently emerged as important regulators in cancer
biology, including ovarian cancer. Owing to their covalently closed-loop structure,
circRNAs exhibit resistance to exonuclease degradation, enabling their stable presence in
cells and body fluids. Accumulating studies indicate that circRNAs contribute to OC
progression by modulating essential cellular processes such as proliferation, migration,
invasion, and apoptosis. Mechanistically, many circRNAs function as competing
endogenous RNAs (ceRNAs), sponging microRNAs and thereby regulating downstream
oncogenes or tumor suppressor genes. Through these interactions, circRNAs influence
key signaling pathways involved in tumor growth and [Link] their functional
roles, circRNAs demonstrate significant clinical potential as diagnostic and prognostic
biomarkers for ovarian cancer. Their differential expression patterns in tumor tissues and
circulation correlate with disease stage, metastasis, and patient survival outcomes.
Furthermore, the stability and detectability of circRNAs in minimally invasive samples
enhance their suitability for liquid biopsy approaches. Collectively, recent findings
highlight the multifaceted roles of circRNAs in ovarian cancer and underscore their
promise as biomarkers and therapeutic targets, offering new avenues for improving OC
management and patient prognosis.

Zhang et al., 2018 stated that Circular RNAs (circRNAs) are a distinct class of
endogenous noncoding RNAs characterized by a covalently closed circular structure that
lacks both 5′ caps and polyadenylated tails. This unique configuration confers exceptional
resistance to exonuclease-mediated degradation, making circRNAs significantly more
stable than their linear counterparts. Advances in high-throughput sequencing
technologies have expanded the understanding of circRNA biogenesis, which primarily
occurs through back-splicing events during pre-mRNA processing. CircRNAs are widely
expressed across eukaryotic cells and often display tissue-specific and developmental
stage–specific expression patterns, suggesting tightly regulated functions in cellular
[Link], circRNAs have emerged as important regulators of gene
expression through multiple mechanisms. One of their most extensively studied roles is
acting as microRNA (miRNA) sponges, thereby modulating post-transcriptional
regulation of target genes. In addition, circRNAs can interact with RNA-binding proteins
(RBPs), influencing protein activity, RNA stability, and transcriptional processes. Some
circRNAs have also been reported to participate in translational regulation, either by
modulating translation of parental genes or, in certain cases, by encoding functional
peptides. These diverse mechanisms position circRNAs as integral components of
complex gene regulatory networks and explain their growing recognition in disease
[Link] to their inherent stability, evolutionary conservation, and abundance in
 28

body fluids such as blood and saliva, circRNAs are increasingly considered promising
diagnostic and prognostic biomarkers for various diseases. Numerous studies have
demonstrated disease-specific circRNA expression profiles, particularly in cancers and
other chronic conditions. However, despite their potential, challenges remain, including
standardization of detection methods, validation in large clinical cohorts, and
understanding context-dependent functions. Overall, current literature underscores both
the advantages and limitations of circRNAs as novel biomarkers, highlighting their
promising but still evolving role in clinical diagnostics and translational medicine.

Wang et al., 2021 determinted that Cancer remains one of the leading causes of mortality
worldwide, largely due to late-stage diagnosis and limited effectiveness of treatment in
advanced disease. Although conventional blood-based biomarkers such as CEA, AFP,
and CA-125 are routinely used in clinical practice, their limited sensitivity and specificity
reduce their effectiveness for early cancer screening in the general population. These
limitations have prompted increasing interest in identifying novel, minimally invasive
diagnostic approaches. Liquid biopsy has emerged as a promising strategy, enabling the
detection of tumor-derived components in body fluids such as blood and urine. Within
this context, circular RNAs (circRNAs) have gained considerable attention as potential
next-generation [Link] are a recently identified class of noncoding RNAs
produced primarily through back-splicing of precursor mRNAs. Advances in high-
throughput transcriptome sequencing have revealed that circRNAs are abundant, stable,
and widely distributed in various body fluids, including plasma, serum, exosomes, and
urine. Their covalently closed structure protects them from exonuclease degradation,
allowing them to persist in extracellular environments. Importantly, accumulating
evidence indicates that circRNA expression profiles differ significantly between cancer
patients and healthy individuals. These distinct expression patterns suggest that
circulating circRNAs may reflect tumor presence, progression, and molecular
characteristics, supporting their utility in cancer detection and [Link]-cancer
analyses have further demonstrated that specific circRNAs are differentially expressed
across multiple tumor types, highlighting their broad clinical relevance. In liquid biopsy
applications, circRNAs have shown potential not only for early diagnosis but also for
prognosis evaluation and disease surveillance. Their detectability in minimally invasive
samples enhances patient compliance and allows for dynamic monitoring of tumor
evolution. Overall, current research underscores the clinical promise of circRNAs in
liquid biopsy-based cancer diagnostics, positioning them as innovative and potentially
more accurate biomarkers for early detection and personalized cancer management.

Brown et al., 2020 experimented that Circular RNAs (circRNAs) are covalently closed
RNA molecules increasingly recognized for their involvement in cancer biology.
Advances in high-throughput sequencing technologies have enabled the identification of
numerous cancer-specific and tissue-specific circRNAs, many of which are cataloged in
publicly accessible databases. These discoveries have expanded understanding of
circRNA diversity and expression patterns across different malignancies. The
conservation of circRNAs across species, along with their abundant and stable expression
in various biological samples, suggests that they are not random by-products of RNA
splicing but functionally relevant regulatory [Link] possess several
 29

characteristics that make them attractive candidates as biomarkers. Their circular

structure confers resistance to exonuclease degradation, allowing them to remain stable in
body fluids such as plasma, saliva, and urine. This stability enhances their suitability for
minimally invasive diagnostic approaches. Numerous studies have reported circRNAs
with diagnostic, predictive, and prognostic significance in various cancers. In some cases,
mechanistic investigations have demonstrated that circRNAs influence tumor
development through microRNA sponging, interaction with RNA-binding proteins, and
modulation of gene expression pathways associated with proliferation, invasion, and
[Link] promising findings, the clinical application of circRNAs as
noninvasive biomarkers for early cancer detection requires further validation. Large-
scale, multicenter studies are necessary to confirm their sensitivity, specificity, and
reproducibility across diverse patient populations. Additionally, standardized detection
methods and improved understanding of circRNA functional mechanisms will be
essential before routine clinical implementation. Overall, current literature highlights
circRNAs as highly promising molecular tools in oncology, with substantial potential for
improving early diagnosis and personalized cancer management.

Ng et al., 2018 investigated that Circular RNAs (circRNAs) constitute a large and
diverse class of endogenously expressed non-coding RNAs generated through back-
splicing events that create covalently closed-loop structures. With the advancement of
high-throughput sequencing technologies, thousands of circRNAs have been identified in
eukaryotic cells, many of which display high sequence conservation and cell type–
specific expression patterns. These findings indicate that circRNAs are evolutionarily
preserved and biologically regulated molecules rather than random splicing artifacts.
Their widespread distribution and regulated expression profiles suggest important roles in
maintaining cellular homeostasis and gene regulatory [Link], circRNAs
participate in a variety of molecular mechanisms that influence cellular physiology and
tumor biology. They are widely recognized for their ability to act as microRNA (miRNA)
sponges, thereby modulating post-transcriptional gene regulation. Additionally,
circRNAs can function as transcriptional regulators, interact with RNA-binding proteins
(RBPs), serve as templates for protein translation, and contribute to immune regulation.
In cancer, circRNA expression patterns have been shown to correlate with
clinicopathological features such as tumor stage, metastasis, and patient survival in
malignancies including breast, liver, gastric, colorectal, and lung cancers. These
mechanistic and clinical associations highlight the involvement of circRNAs in tumor
initiation, progression, and [Link] a translational perspective, circRNAs possess
several characteristics that make them promising molecular biomarkers. Their high
stability, resistance to exonuclease degradation, and presence in accessible body fluids
such as blood and saliva support their potential application in non-invasive diagnostic and
monitoring strategies. Moreover, circRNAs may serve as prognostic indicators and even
therapeutic targets in cancer management. However, further research is necessary to fully
elucidate the molecular interactions and signaling pathways mediated by circRNAs in
tumors. A deeper understanding of these mechanisms will be essential for advancing
circRNA-based applications in clinical oncology and personalized medicine.
 30

Xia et al., 2018 demonstrated that Circular RNAs (circRNAs) are a novel class of
endogenous noncoding RNAs that differ structurally and functionally from linear RNAs
due to their covalently closed-loop configuration. This unique structure lacks 5′ and 3′
ends, rendering circRNAs resistant to exonuclease-mediated degradation, particularly by
RNase R. As a result, circRNAs exhibit remarkable stability and persistence in eukaryotic
cells and body fluids. High-throughput sequencing studies have demonstrated that
circRNAs are abundantly expressed, evolutionarily conserved, and often display
developmental stage–specific and tissue-specific expression patterns. These intrinsic
characteristics distinguish circRNAs from other noncoding RNAs and underscore their
biological significance.

Accumulating evidence indicates that circRNAs are closely associated with the
pathogenesis of numerous diseases, especially cancer. Aberrant circRNA expression
profiles have been identified across various tumor types, suggesting their involvement in
oncogenic transformation and tumor progression. Mechanistically, circRNAs can regulate
gene expression by targeting key oncogenes or tumor suppressor genes, frequently
through interactions with microRNAs and RNA-binding proteins. By participating in
regulatory networks that control proliferation, apoptosis, migration, and metastasis,
circRNAs contribute to critical steps in carcinogenesis. Their dysregulation often
correlates with tumor stage, metastasis, and patient prognosis, highlighting their
functional and clinical [Link], circRNAs are not confined to intracellular
environments; they can be released from tissues and cells into extracellular fluids such as
blood and saliva. Their abundance and stability in these accessible body fluids make them
highly attractive candidates for noninvasive diagnostic and prognostic biomarkers. Given
their specificity and detectability, circRNAs hold substantial promise for improving early
cancer detection and monitoring disease progression. Overall, recent research emphasizes
the expanding role of circRNAs in cancer biology and supports further investigation into
their potential application as reliable biomarkers in clinical oncology.

Chaichian et al., 2020 described that Advances in RNA sequencing (RNA-Seq) and
biochemical analyses have led to the identification of circular RNAs (circRNAs) as a
distinct class of RNA molecules generated through alternative splicing events. Unlike
linear messenger RNAs, circRNAs form covalently closed-loop structures that contribute
to their remarkable stability within cells. Studies have shown that circRNAs are
abundantly expressed and often display cell-specific, tissue-specific, and developmental
stage–specific expression patterns, suggesting tightly regulated biological functions.
These characteristics indicate that circRNAs possess significant regulatory potential and
are actively involved in diverse cellular processes rather than being mere splicing by-
[Link] recent years, circRNAs have attracted considerable research interest due to
their functional roles in gene regulation and disease progression. Accumulating evidence
demonstrates that circRNAs participate in multiple cancers by modulating key molecular
pathways involved in cell proliferation, apoptosis, invasion, and metastasis. Their ability
to act as microRNA sponges, interact with RNA-binding proteins, and influence
transcriptional regulation highlights their importance in tumor biology. Moreover, the
stability and detectability of circRNAs in tissues and body fluids position them as
promising biomarkers for cancer diagnosis and [Link] cancer, a leading
 31

cause of cancer-related mortality among women worldwide, has become a significant

focus of circRNA research. Emerging studies have identified aberrant circRNA
expression profiles in cervical cancer tissues, implicating them in carcinogenesis and
tumor progression. Understanding the molecular mechanisms by which circRNAs
contribute to cervical neoplasia may facilitate the development of novel transcription-
based therapeutic strategies. Overall, current literature underscores the growing
recognition of circRNAs in cervical cancer biology and highlights their potential as
diagnostic biomarkers and therapeutic targets in future clinical applications.

Lü et al., 2017 revealed that Circular RNAs (circRNAs) have increasingly been
recognized as important regulators in cancer biology; however, their comprehensive
expression patterns in breast cancer were not fully understood until large-scale profiling
studies were conducted. Using high-throughput microarray analysis, researchers
constructed a genome-wide circRNA expression profile to compare breast cancer tissues
with normal controls. This systematic approach identified 1,155 differentially expressed
circRNAs, of which 715 were upregulated and 440 were downregulated in tumor
samples. These findings highlight the widespread dysregulation of circRNAs in breast
cancer and suggest that circRNA alterations are closely associated with tumor
development and [Link] validate the microarray data, selected circRNAs were
examined using quantitative real-time polymerase chain reaction (qRT-PCR), confirming
consistent differential expression patterns. Specifically, hsa_circ_103110,
hsa_circ_104689, and hsa_circ_104821 were significantly upregulated, whereas
hsa_circ_006054, hsa_circ_100219, and hsa_circ_406697 were downregulated in breast
cancer tissues. Bioinformatic and sequence analyses further predicted that these
circRNAs contain complementary microRNA response elements, indicating their
potential role as competing endogenous RNAs (ceRNAs). Through miRNA
sequestration, these circRNAs may regulate downstream oncogenic or tumor-suppressive
pathways, thereby contributing to [Link], the study also evaluated
the diagnostic performance of selected circRNAs. A combined panel of three
downregulated circRNAs (hsa_circ_006054, hsa_circ_100219, and hsa_circ_406697)
demonstrated a strong discriminatory ability for distinguishing breast cancer tissues, with
an area under the receiver operating characteristic (ROC) curve of 0.82. This level of
diagnostic accuracy underscores the potential clinical utility of circRNAs as biomarkers
for breast cancer detection. Overall, the findings provide compelling evidence that
circRNAs are significantly dysregulated in breast cancer and play functional roles in
tumor-related pathways, reinforcing their promise as diagnostic indicators and
contributors to cancer pathogenesis.

Zhang et al., 2017 examined that In recent years, circular RNAs (circRNAs) have
emerged as important regulatory molecules in cancer biology, attracting significant
scientific interest due to their diverse functional roles. Their unique covalently closed-
loop structure provides enhanced stability compared to linear RNAs, enabling them to
persist in both intracellular and extracellular environments. Increasing evidence indicates
that circRNAs are involved in multiple aspects of tumor biology, including cell
proliferation, apoptosis, invasion, and metastasis. Their ability to regulate gene
expression through mechanisms such as microRNA (miRNA) sponging, interaction with
 32

RNA-binding proteins, and modulation of transcriptional activity highlights their

significance in oncogenic processes.

Beyond their biological roles, circRNAs have shown great promise in the field of
personalized or individualized cancer medicine. Due to their tissue-specific expression
patterns and detectability in body fluids, circRNAs can serve as reliable biomarkers for
cancer diagnosis, prognosis, and disease monitoring. Additionally, their involvement in
key signaling pathways makes them attractive therapeutic targets. Strategies such as
silencing oncogenic circRNAs or restoring tumor-suppressive circRNAs are being
explored as potential approaches for targeted cancer therapy. These developments
suggest that circRNAs could play a crucial role in improving precision medicine and
tailoring treatment strategies based on individual molecular profiles.

Despite these promising prospects, several challenges hinder the clinical translation of
circRNA-based applications. These include the need for standardized detection methods,
validation in large-scale clinical studies, and a deeper understanding of their complex
regulatory mechanisms. Furthermore, efficient and safe delivery systems for circRNA-
targeted therapies remain to be developed. Nevertheless, ongoing research continues to
expand our understanding of circRNAs, offering new insights into cancer molecular
biology. Overall, circRNAs represent a promising frontier in oncology, with the potential
to enhance diagnostic accuracy and open new avenues for innovative cancer treatment
strategies.

Han et al., 2017 proclaimed that Circular RNAs (circRNAs) represent a novel and
diverse class of endogenous noncoding RNAs characterized by a covalently closed-loop
structure that lacks 5′ and 3′ ends. Unlike linear RNAs, this circular configuration
provides resistance to exonuclease-mediated degradation, contributing to their
remarkable stability within cells. CircRNAs are widely expressed in eukaryotic
organisms and are often abundant, evolutionarily conserved, and tissue- or developmental
stage–specific in their expression. They are primarily generated through back-splicing
events from exonic or intronic sequences, including lariat-driven circularization. These
structural and biogenetic features underscore their regulated formation and functional
relevance in cellular [Link], circRNAs participate in multiple regulatory
mechanisms that influence gene expression and cellular homeostasis. One of their most
extensively studied roles is acting as microRNA (miRNA) sponges, whereby they bind
and sequester miRNAs to modulate downstream target gene expression. In addition,
circRNAs can regulate gene transcription and expression by interacting with
transcriptional machinery or serving as RNA-binding protein (RBP) sponges, thereby
influencing protein function and RNA metabolism. Emerging studies also suggest that
certain circRNAs possess protein-coding potential, expanding their functional versatility.
Through these mechanisms, circRNAs are involved in critical biological pathways that
contribute to tumor initiation, progression, and [Link] evidence highlights
the clinical significance of circRNAs in cancer diagnosis and therapy. Aberrant circRNA
expression profiles have been identified across various malignancies, suggesting their
potential utility as novel biomarkers and therapeutic targets. Advances in circRNA
research methodologies, including RNA sequencing, microarray analysis, and
 33

quantitative PCR, have facilitated the detection and functional characterization of

circRNAs in both tissues and body fluids. Collectively, current literature emphasizes the
importance of understanding circRNA origins, characteristics, and molecular functions,
as this knowledge may support the development of circRNA-based diagnostic tools and
targeted therapeutic strategies in oncology.

Garlapati et al., 2021 studied that Circular RNAs (circRNAs) have emerged as important
regulatory molecules implicated in a wide range of pathological processes, particularly
cancer. The advent of high-throughput RNA sequencing technologies has led to the
identification of numerous circRNAs that are differentially expressed across various
cancer cell lines and tumor tissues. These findings have challenged the earlier perception
of circRNAs as splicing by-products and instead established them as functionally
significant components of gene regulatory networks. Differential circRNA expression
patterns observed in malignancies suggest their involvement in tumor initiation,
progression, and [Link], circRNAs regulate cell signaling pathways
by modulating gene expression at transcriptional and post-transcriptional levels. Many
circRNAs function as microRNA (miRNA) sponges, thereby influencing the expression
of downstream target genes involved in proliferation, apoptosis, invasion, and
angiogenesis. Others interact with RNA-binding proteins or participate in the regulation
of transcriptional machinery, further contributing to alterations in cellular signaling
cascades. Through these mechanisms, circRNAs can either promote oncogenic pathways
or act as tumor suppressors, depending on their specific molecular interactions and cancer
[Link] the distribution of circRNAs across different cancer types and
elucidating their mechanisms of action is essential for advancing translational research.
By mapping circRNA-mediated regulatory networks, researchers can identify potential
diagnostic and prognostic biomarkers that reflect tumor-specific molecular alterations.
Additionally, targeting dysregulated circRNAs offers a promising therapeutic strategy for
cancer treatment. Overall, the expanding body of research underscores the importance of
circRNAs in cancer biology and highlights their potential as novel tools for improving
cancer diagnosis, prognosis, and targeted therapy.

Tang et al., 2020 stated that Circular RNAs (circRNAs) represent an intriguing and
recently recognized class of noncoding RNAs characterized by a covalently closed-loop
structure that confers exceptional stability and resistance to exonuclease-mediated
degradation. As emerging members of the ncRNA family, circRNAs have attracted
growing scientific attention due to their conserved nature, abundance, and tissue- and
developmental stage–specific expression patterns. High-throughput transcriptomic
analyses have identified more than 30,000 circRNAs across eukaryotic systems,
highlighting their widespread distribution and biological relevance. Their structural
uniqueness allows them to maintain stability more effectively than linear RNAs,
reinforcing their potential functional importance in cellular [Link]
evidence indicates that circRNAs are actively involved in multiple hallmarks of cancer,
including tumor initiation, growth, invasion, metastasis, apoptosis, and angiogenesis.
Dysregulated circRNA expression has been observed in a wide range of malignancies,
suggesting their contribution to cancer pathogenesis. Functionally, circRNAs exert
regulatory effects through diverse molecular mechanisms such as acting as microRNA
 34

(miRNA) sponges, functioning as protein scaffolds, interacting with RNA-binding

proteins (RBPs), and regulating autophagy-related pathways. Through these interactions,
circRNAs influence key signaling cascades that govern cancer cell behavior, thereby
contributing to both tumor promotion and suppression depending on their specific
[Link] a clinical perspective, circRNAs have emerged as promising candidates for
cancer prevention, diagnosis, prognosis, and therapeutic targeting. Their stable presence
in tissues and body fluids, along with their disease-specific expression patterns, makes
them highly suitable as non-invasive biomarkers. Furthermore, circRNAs have been
implicated in drug resistance mechanisms and regulatory signaling pathways,
underscoring their potential relevance in personalized cancer therapy. Overall, current
literature strongly supports the significance of circRNAs in cancer biology, emphasizing
their expanding role as diagnostic tools and therapeutic targets in oncology research and
clinical applications.

Dong et al., 2017 determined that Circular RNAs (circRNAs) are a class of endogenous
RNAs characterized by a covalently closed continuous loop structure, which
distinguishes them from linear RNAs. They are widely expressed in mammalian cells and
are highly conserved across species, often exhibiting tissue- and developmental stage–
specific expression patterns. Initially considered by-products of splicing errors, circRNAs
were long overlooked in molecular biology. However, advances in high-throughput
sequencing and bioinformatics analyses have revealed that circRNAs are stable,
abundant, and functionally significant molecules with regulated biogenesis and
[Link] research has demonstrated that circRNAs perform diverse
regulatory functions within the cell. One of their most well-characterized roles is acting
as microRNA (miRNA) sponges, thereby modulating gene expression by sequestering
miRNAs and preventing them from binding to their target mRNAs. In addition to miRNA
sponging, circRNAs can regulate transcription and alternative splicing, interact with
RNA-binding proteins, and influence gene expression at multiple levels. These functional
capabilities highlight their importance in maintaining cellular homeostasis and regulating
key biological [Link] evidence strongly associates circRNAs with various
human diseases, particularly cancer. Dysregulated circRNA expression has been
observed in numerous malignancies, where they contribute to tumorigenesis, progression,
and metastasis. Due to their stability, specificity, and detectability in body fluids,
circRNAs are increasingly recognized as promising diagnostic and prognostic
biomarkers. Following the discovery of microRNAs and long noncoding RNAs
(lncRNAs), circRNAs have now become a major focus in cancer-related RNA research.
Overall, current literature underscores their significant biological roles and potential
clinical applications in oncology.

Tian et al., 2021 experimented that Triple-negative breast cancer (TNBC) is recognized
as the most aggressive molecular subtype of breast cancer, accounting for approximately
10–20% of all cases. It is characterized by the absence of estrogen receptor (ER),
progesterone receptor (PR), and HER2 expression, which limits the availability of
targeted hormonal or HER2-directed therapies. TNBC occurs more frequently in younger
women and is associated with high proliferative capacity, early metastasis, and poor
clinical prognosis. Although chemotherapy remains the primary systemic treatment
 35

option, therapeutic resistance and high recurrence rates highlight the urgent need to
identify novel and precise molecular targets to improve patient [Link] recent years,
circular RNAs (circRNAs), a class of non-coding RNAs abundantly expressed in
eukaryotic cells, have emerged as important regulators in cancer biology. CircRNAs
exert diverse cellular functions, including acting as microRNA (miRNA) sponges,
interacting with RNA-binding proteins (RBPs), regulating gene transcription, and in
some cases serving as templates for translation. Increasing evidence suggests that
dysregulated circRNAs are involved in TNBC tumorigenesis, proliferation, invasion,
metastasis, and chemoresistance. Through modulation of key oncogenic signaling
pathways, circRNAs may function either as tumor promoters or suppressors, thereby
influencing disease progression and therapeutic [Link] their stability, tissue
specificity, and detectability in body fluids, circRNAs hold considerable promise as
diagnostic and prognostic biomarkers for TNBC. Moreover, their regulatory roles in gene
expression and cancer-associated pathways suggest potential utility as therapeutic targets.
However, despite encouraging findings, the clinical application of TNBC-associated
circRNAs remains limited by insufficient functional validation and lack of large-scale
clinical studies. Future research focusing on mechanistic elucidation, standardized
detection methods, and translational studies will be essential to facilitate the integration
of circRNA-based strategies into clinical management of TNBC.

Verduci et al., 2021 investigated that Circular RNAs (circRNAs) are a distinct class of
endogenous RNAs characterized by a covalently closed-loop structure that confers
exceptional stability compared to linear RNAs. Although their overall abundance is
relatively low, their circular configuration protects them from exonuclease degradation,
enabling prolonged persistence within cells. CircRNAs also exhibit a high degree of
tissue specificity, suggesting tightly regulated expression patterns and potential functional
importance in maintaining cellular homeostasis. Advances in molecular biology have
clarified their biogenesis, primarily through back-splicing events, and have shifted the
perception of circRNAs from splicing by-products to biologically active regulatory
[Link], the most well-established role of circRNAs is their activity as
microRNA (miRNA) sponges, whereby they sequester miRNAs and modulate
downstream gene expression. However, emerging evidence indicates that circRNAs
possess additional mechanisms of action, including binding to specific RNA-binding
proteins (RBPs) and, in some cases, serving as templates for the translation of short
peptides. These multifunctional properties enable circRNAs to participate in diverse
molecular signaling pathways. Dysregulated circRNA expression has been implicated in
numerous human diseases, particularly organ-specific cancers such as head and neck,
breast, and lung cancers. Beyond oncology, circRNAs have also been associated with
metabolic disorders like diabetes, as well as cardiovascular and neurodegenerative
diseases, highlighting their broad pathological [Link] growing evidence of
their involvement in various signaling networks, differing and sometimes controversial
interpretations of circRNA functions remain. The complex interplay between circRNAs,
other non-coding RNAs, and RNA-binding proteins suggests that they contribute to
intricate regulatory networks that maintain cellular equilibrium. Given their remarkable
stability and tissue specificity, circRNAs are increasingly considered promising
biomarkers for disease diagnosis, prognosis, and monitoring of treatment response.
 36

Continued research is essential to resolve existing uncertainties and to fully harness their
potential in clinical applications.

Song et al., 2024 demonstrated that Infertility affects approximately 10% of individuals
during their reproductive years and remains a significant global health concern. Despite
advances in assisted reproductive technologies (ART), including in vitro fertilization
(IVF), implantation failure and adverse pregnancy outcomes continue to limit therapeutic
success. These challenges are often associated with altered gene expression patterns
occurring during both in vivo and in vitro fertilization processes. In recent years, circular
RNAs (circRNAs) have emerged as important epigenetic regulators involved in
reproductive physiology and pathology. Their stable, covalently closed-loop structure and
regulatory capacity position them as key modulators of gene expression in fertility-
related [Link] participate in numerous cellular functions relevant to
reproduction, including nuclear transport, X-chromosome inactivation, apoptosis, stem
cell differentiation, and genomic imprinting. Advances in high-throughput sequencing
technologies have led to the identification of numerous circRNAs with critical roles in
male reproductive biology, such as promoting testicular development, maintaining
spermatogonial stem cell self-renewal and differentiation, and regulating spermatocyte
meiosis. In the female reproductive system, circRNAs have been implicated in ovarian
function, embryo implantation, pregnancy-related disorders, endometriosis, and
gynecological cancers. Their involvement in these processes highlights their regulatory
influence across multiple stages of reproductive development and [Link] their
stability, tissue specificity, and detectability in biological fluids, circRNAs hold
significant promise as diagnostic biomarkers and therapeutic targets for reproductive
disorders in both males and females. Particularly in women’s infertility, circRNA-
mediated regulatory networks may provide novel insights into the molecular mechanisms
underlying implantation failure and other reproductive complications. Comprehensive
investigation of circRNA expression profiles and functional roles will be essential for
translating experimental findings into clinical applications aimed at improving fertility
outcomes.

De Palma et al., 2022 reported that breast cancer (BC) is a highly heterogeneous disease
that often remains asymptomatic in its early stages, making early diagnosis and effective
management challenging. Current clinical strategies rely heavily on imaging techniques
combined with immune histochemical and molecular evaluations to enable early
detection, patient stratification, and personalized treatment planning. These diagnostic
approaches assist clinicians in determining prognosis and selecting appropriate
therapeutic regimens. However, the complexity and variability of BC highlight the need
for more precise and reliable molecular biomarkers to improve clinical outcomes.
Circular RNAs (circRNAs), a recently recognized class of long non-coding RNAs, have
emerged as significant regulators in breast carcinogenesis and tumor progression.
Growing evidence suggests that dysregulated circRNAs contribute to key oncogenic
processes, including cell proliferation, invasion, metastasis, and drug resistance.
Additionally, specific circRNA expression patterns have been correlated with clinic
pathological characteristics of breast cancer patients, underscoring their potential role in
disease classification and outcome prediction. Mechanistically, circRNAs regulate gene
 37

expression through interactions with microRNAs, RNA-binding proteins, and signaling

pathways involved in cancer [Link] to their covalently closed-loop structure,
circRNAs exhibit remarkable stability and resistance to degradation, allowing their
detection in various body fluids such as blood and plasma. This stability, combined with
tissue-specific expression patterns, makes them promising non-invasive biomarkers for
diagnosis, prognosis, and therapeutic monitoring in breast cancer. Collectively, current
literature highlights the clinical relevance of circRNAs and supports their potential
integration into molecular diagnostic and personalized treatment strategies for breast
cancer management.

Tang et al., 2020 described that Circular RNAs (circRNAs) are a unique class of
endogenous non-coding RNAs characterized by a covalently closed-loop structure that
provides remarkable stability and resistance to exonuclease degradation. Unlike linear
RNAs, circRNAs lack 5′ and 3′ ends, which contributes to their conserved nature and
sustained expression in various tissues. With the advancement of high-throughput
sequencing technologies, more than 30,000 circRNAs have been identified, many of
which display tissue- and developmental stage–specific expression patterns. These
characteristics, along with their abundance and evolutionary conservation, have
positioned circRNAs as significant regulatory molecules rather than mere by-products of
aberrant [Link] evidence demonstrates that dysregulated circRNAs are
involved in multiple aspects of cancer biology, including tumorigenesis, proliferation,
invasion, metastasis, apoptosis, and angiogenesis. Functionally, circRNAs exert their
effects through diverse mechanisms such as acting as microRNA (miRNA) sponges,
serving as protein scaffolds, regulating autophagy, and interacting with RNA-binding
proteins (RBPs). Through these interactions, circRNAs influence critical oncogenic
signaling pathways and gene regulatory networks, contributing to both tumor-promoting
and tumor-suppressive processes depending on the cellular context. Their involvement in
drug resistance and signaling pathway modulation further emphasizes their importance in
cancer [Link] their high stability, specificity, and detectability in tissues and
body fluids, circRNAs hold considerable promise as diagnostic and prognostic
biomarkers, as well as potential therapeutic targets in oncology. Understanding their
expression profiles, regulatory mechanisms, and roles in different cancer types provides
valuable insights into disease pathogenesis and treatment strategies. Overall, current
literature strongly supports the clinical relevance of circRNAs and highlights their
potential utility in improving cancer diagnosis, prognosis, and targeted therapy.

Tang et al., 2021 revealed that Colorectal cancer (CRC) remains one of the most
prevalent malignancies worldwide and is currently ranked among the leading causes of
cancer-related mortality. Over the past two decades, its incidence has increased
significantly, with a concerning rise among individuals unde the age of 50. Despite
improvements in surgical techniques, chemotherapy, radiotherapy, and targeted
molecular therapies, the overall survival rate of CRC patients remains unsatisfactory,
largely due to late-stage diagnosis and disease progression. These limitations have driven
growing interest in understanding the molecular mechanisms underlying CRC
development, particularly the role of noncoding RNAs (ncRNAs) in tumor
[Link] RNAs (circRNAs), a subclass of endogenous ncRNAs characterized by
 38

a covalently closed-loop structure, have emerged as important regulatory molecules in

CRC. Their unique structure confers exceptional stability compared to linear RNAs,
enhancing their suitability for clinical applications. Numerous studies have demonstrated
that circRNAs are differentially expressed in CRC tissues and may function either as
oncogenes or tumor suppressors. Mechanistically, circRNAs can act as microRNA
(miRNA) sponges, regulate mRNA expression, and modulate key oncogenic and tumor-
suppressive signaling pathways. Through these interactions, circRNAs influence CRC
initiation, progression, metastasis, and therapeutic [Link] their stability, cancer-
specific expression patterns, and involvement in critical molecular pathways, circRNAs
hold substantial promise as diagnostic, prognostic, and therapeutic biomarkers in CRC.
Their potential utility in early detection could address one of the major challenges in
CRC management—late diagnosis—while their regulatory roles offer novel opportunities
for targeted therapeutic intervention. However, further large-scale clinical studies are
required to validate their effectiveness and facilitate the translation of experimental
findings into routine clinical practice. Overall, current literature underscores the
transformative potential of circRNAs in improving CRC diagnosis, prognosis, and
treatment strategies.

Su et al., 2019 reported that circular RNAs (circRNAs) are a distinct class of RNA
molecules characterized by their covalently closed-loop structure, which confers
remarkable stability compared to linear RNAs. They are abundantly expressed in
eukaryotic cells and display location- and developmental stage–specific expression
patterns, indicating tightly regulated biogenesis and functional relevance. Advances in
RNA sequencing technologies have significantly expanded the identification and
characterization of circRNAs, leading to growing recognition of their regulatory roles in
gene expression. Rather than being considered splicing by-products, circRNAs are now
understood to be active participants in diverse molecular [Link],
circRNAs contribute to gene regulation through multiple mechanisms. The most
extensively studied role involves acting as microRNA (miRNA) sponges, thereby
modulating post-transcriptional gene expression. Additionally, circRNAs can interact
with RNA-binding proteins, regulate transcription and alternative splicing, and in certain
cases serve as templates for protein translation. These versatile functions enable
circRNAs to influence critical cellular pathways. Emerging evidence highlights their
involvement in various hallmarks of cancer, including sustained proliferation, evasion of
apoptosis, metastasis, angiogenesis, and therapeutic resistance. Moreover, circRNAs have
been implicated in maintaining cancer stemness, further underscoring their role in tumor
progression and [Link] their stability, specificity, and functional diversity,
circRNAs have garnered attention as promising biomarkers for cancer diagnosis and
prognosis. Their presence in body fluids and tumor tissues enhances their potential utility
in non-invasive detection and monitoring of disease progression. Expanding knowledge
of circRNA biology not only deepens understanding of cancer molecular mechanisms but
also opens new avenues for targeted therapeutic strategies. Overall, current research
supports the significance of circRNAs in cancer development and highlights their
emerging role in precision oncology.
 39

Sufianov et al., 2023 demonstrated that lung cancer remains the leading cause of cancer-
related mortality worldwide, contributing significantly to global morbidity and healthcare
burden. Although cigarette smoking is the primary risk factor, a considerable proportion
of lung cancer cases occur in non-smokers, indicating the involvement of complex
genetic and epigenetic mechanisms in disease pathogenesis. Despite advances in imaging
techniques, surgical approaches, chemotherapy, radiotherapy, and targeted therapies, the
overall prognosis of lung cancer patients remains poor due to late diagnosis and
therapeutic resistance. These challenges highlight the urgent need for reliable molecular
markers and effective therapeutic targets to improve early detection and patient
outcomes. Circular RNAs (circRNAs), a class of covalently closed non-coding RNAs,
have emerged as important regulators in lung cancer biology. Their structural stability,
evolutionary conservation, and tissue-specific expression patterns make them attractive
candidates for clinical applications. Numerous studies have demonstrated that circRNAs
are dysregulated in lung cancer and contribute to tumor progression through diverse
mechanisms, particularly by acting as microRNA (miRNA) sponges. By modulating
miRNA activity and downstream gene expression, circRNAs influence key oncogenic
pathways involved in cell proliferation, migration, invasion, and metastasis. Recent
research increasingly supports the clinical relevance of circRNAs in lung cancer
diagnosis, treatment, and prognosis. Differential expression profiles of circRNAs in
tumor tissues and body fluids suggest their potential utility as non-invasive diagnostic
and prognostic biomarkers. Furthermore, targeting circRNA-mediated regulatory
networks may offer novel therapeutic strategies. Overall, current evidence underscores
the significance of circRNAs in lung cancer pathogenesis and highlights their promising
role in advancing molecular diagnosis and precision therapy.

Huang et al., 2021 stated that lung cancer continues to exhibit high incidence and
mortality rates worldwide, making early detection a critical factor in improving patient
survival. Currently, diagnosis primarily relies on imaging techniques and tissue biopsy;
however, these approaches present several limitations, including low specificity,
invasiveness, and challenges associated with repeated sampling. Such constraints reduce
their effectiveness in early-stage detection and disease monitoring. Consequently, there is
an urgent need for more sensitive, specific, and minimally invasive biomarkers that can
enhance early diagnosis and improve clinical outcomes, particularly in non-small cell
lung cancer (NSCLC), the most common subtype of lung [Link] studies have
highlighted circular RNAs (circRNAs) as key regulatory molecules involved in cancer
progression and potential biomarkers for various malignancies. CircRNAs are
characterized by their covalently closed-loop structure, which confers remarkable
stability and resistance to degradation. In NSCLC, numerous circRNAs have been found
to be aberrantly expressed and to participate in tumorigenic processes such as
proliferation, invasion, metastasis, and therapeutic resistance. Their disease-specific
expression patterns and functional involvement in oncogenic pathways make them
promising candidates for diagnostic and prognostic [Link], circRNAs
are stably present in exosomes and circulating body fluids, positioning them as ideal
targets for liquid biopsy approaches. Liquid biopsy offers significant advantages,
including high sensitivity, specificity, and the ability to perform repeated sampling for
real-time disease monitoring. The detectability of circRNAs in circulating nucleic acids
 40

further enhances their clinical value as non-invasive biomarkers for NSCLC. Overall,
accumulating evidence supports the potential of circRNAs to transform early detection
strategies and improve diagnostic accuracy in NSCLC, although further validation
through large-scale clinical studies remains essential.