Diagnostic and prognostic significance of internal tRNA-derived fragment glycine-glycine-cytosine (itRNA^GlyGCC) expression in epithelial ovarian cancer

1. Introduction

Epithelial ovarian cancer (EOC) was estimated to account for 4.7% of cancer deaths and 3.7% of cases worldwide, making it the eighth most prevalent cancer in women [1]. The two primary characteristics of EOC are its insidious progression and late-stage diagnosis; early and accurate identification of the disease is crucial for improving survival rates [2]. Approximately 70% of cases of EOC are diagnosed at an advanced stage, resulting in a 5-year survival rate of only 30% for patients. However, if EOC is identified early and confined to the ovaries, the 5-year survival rate could be as high as 90% [3].

The early symptoms of EOC are vague, making it challenging to identify the disease early despite advancements in our understanding of its molecular and cellular basis. This highlights the urgent need for new molecular biomarker-based therapeutic approaches and new diagnostic and prognostic markers.

Among the diverse types of non-coding RNAs (ncRNAs), tRNA-derived RNA fragments (tRFs) have recently emerged as a promising area with significant diagnostic and prognostic potential [4]. These endogenous, single-stranded non-coding RNA fragments typically range from 14 to 40 nucleotides in length and exhibit functional similarities to other ncRNA classes, such as miRNAs, playing a critical role in the regulation of gene expression [5, 6]. Despite ongoing research, the precise biological functions of transfer RNAs (tRNAs) remain incompletely understood. One notable example is the internal tRNA-derived fragment GlyGCC (i-tRF^GlyGCC), which has the nucleotide sequence 5′-GCATTGGTGGTTCAGTGGTAGAATTCTCGC-3′. This 30-nucleotide fragment is derived from the internal regions of mature glycine tRNA containing the GCC anticodon and is generated through specific enzymatic cleavage of the parent tRNA [7].

Emerging evidence suggests that i-tRF^GlyGCC is not merely a byproduct of tRNA degradation but a functionally significant molecule with diverse biological roles. Dysregulation of i-tRF^GlyGCC expression has been observed in various pathological conditions, particularly in cancer development and progression [8]. Mechanistically, i-tRF^GlyGCC has been implicated in interactions with key cellular proteins and the regulation of gene expression through both transcriptional and post-transcriptional mechanisms [9]. Studies have demonstrated its involvement in critical cellular processes, including cell proliferation, apoptosis and stress response pathways [10]. For instance, Yang et al. [11] reported that i-tRF^GlyGCC can modulate protein synthesis by competing with full-length tRNAs. At the same time, Sarkar et al. [12] identified its role in microRNA-like gene silencing through direct interactions with target mRNAs.

The evolutionary conservation of i-tRF^GlyGCC across species, combined with its tissue-specific expression patterns, underscores its fundamental role in cellular regulation. This molecular signature has emerged as a crucial factor in both physiological and pathological processes, particularly in cancer biology [13]. tRNAs demonstrate a dual regulatory function in malignancy, acting as both oncogenic and oncostatic elements through their roles as inducers and blockers of cancer-related pathways [9]. Their tumor-suppressive effects are mediated through post-transcriptional regulation of cancer-associated genes, operating via transcriptional control mechanisms and malignant gene stabilization [14].

Studies have expanded our understanding of tRF involvement in disease pathogenesis. These molecules have been implicated in a broad spectrum of pathological conditions, including hematologic malignancies [15], solid tumors [16], metabolic disorders [17], inflammatory conditions [13], viral infections [18] and neurological diseases [9]. This diverse disease association highlights the expansive regulatory network influenced by tRF expression and function. Of particular significance is tRF-03357, which has demonstrated the capacity to enhance epithelial ovarian cancer (EOC) cell migration, invasion and proliferation in vitro [19]. Furthermore, i-tRFs derived from tRNA-Gly have emerged as promising diagnostic biomarkers for ovarian cancer, exhibiting distinctive expression patterns in patient serum compared to healthy controls [20]. The high specificity and sensitivity of these i-tRFs in ovarian cancer prediction, coupled with their dysregulation in malignancy and prevalence in body fluids, position them as valuable potential molecular markers for EOC diagnosis and monitoring.

This study investigates the potential of i-tRF^GlyGCC as a diagnostic and prognostic molecular marker in EOC.

2. Patients and methods

The study involved 142 patients diagnosed with stage III epithelial ovarian cancer according to the International Federation of Gynecology and Obstetrics (FIGO) staging, between May 2018 and June 2024 [21]. Pathological examination confirmed epithelial ovarian cancer (EOC). Debulking surgery had been performed on each patient within two months before the first course of chemotherapy. One hundred healthy women volunteered to serve as controls for the study.

The inclusions included being 18 or older, having normal baseline blood counts, and demonstrating normal kidney and liver function.

The study excluded those with ovarian low-malignant tumors, a performance status exceeding two on the Eastern Cooperative Oncology Group (ECOG) scale [22], or a glomerular filtration rate (GFR) below 60 mL/minute. Additionally, patients with severe neuropathy, a history of prior chemotherapy or radiotherapy for ovarian cancer, congestive heart failure, or arrhythmias were omitted.

All participants provided written informed consent after Zagazig University’s Research Ethics Committee granted authorization (ZU-232/2018).

After surgery, all patients received a standardized postoperative chemotherapy regimen. This regimen included six cycles; each repeated every three weeks. Each cycle consisted of the following treatments:

- Paclitaxel: administered intravenously at a 175 mg/m² dose over three hours.

- Cisplatin: administered intravenously at a 75 mg/m² dose following adequate patient hydration.

3. Results

3.2 Expression of i-tRF^GlyGCC in epithelial ovarian cancer: a potential biomarker?

The expression of i-tRF^GlyGCC was found to be downregulated in epithelial ovarian cancer. In epithelial ovarian cancer samples, the mean ± SD level of i-tRF^GlyGCC was 5.995 ± 3.302. In contrast, in normal ovarian tissue (ANOT), it was 0.4080 ± 0.1707, indicating a statistically significant difference (p < 0.0001) (Fig. 1A). Similar results were observed in the serum of patients with epithelial ovarian cancer compared to healthy controls. The mean expression of i-tRF^GlyGCC in the serum of ovarian cancer patients was 6.579 ± 2.949, while in the control group, it was 0.3666 ± 0.1905, again demonstrating a statistically significant difference (p < 0.0001) (Fig. 1B).

The effectiveness of i-tRF^GlyGCC as a diagnostic marker for epithelial ovarian cancer (EOC) was evaluated by analyzing its performance using receiver operating characteristic (ROC) curves. The findings revealed exceptional diagnostic accuracy for i-tRF^GlyGCCin EOC tissues (Fig. 1C) and patient serum samples (Fig. 1D). In EOC tissues, i-tRF^GlyGCC perfectly differentiated between malignant and normal ovarian tissues, with an AUC of 1.000 (95% Confidence Interval (CI): 0.9998–1.000; p < 0.0001). Similarly impressive results were observed in the serum analysis, where i-tRF^GlyGCC effectively distinguished EOC patients from controls (AUC = 1.000; 95% CI: 1.000–1.000; p < 0.0001).

Fig. 1.Expression and diagnostic performance of i-tRF^GlyGCC in epithelial ovarian cancer (EOC). (A) i-tRF^GlyGCC expression levels in EOC tissues compared to adjacent normal ovarian tissues (ANOT). (B) i-tRF^GlyGCC expression levels in the serum of EOC patients versus healthy controls. (C) Receiver operating characteristic (ROC) curve analysis of i-tRF^GlyGCC expression in EOC tissues, demonstrating diagnostic accuracy (AUC = 1.000). (D) ROC curve analysis of i-tRF^GlyGCC expression in serum, showing diagnostic accuracy (AUC = 1.000). All comparisons were statistically significant (p < 0.0001). AUC: Area Under Curve; i-tRF^GlyGCC: internal tRNA-derived fragment Glycine-Glycine-Cytosine; CI: Confidence interval.

We studied the relationship between i-tRF^GlyGCC levels in epithelial ovarian cancer (EOC) tissues and patient serum. The results revealed a robust positive correlation, indicating that tissue and serum levels closely mirror each other (R² = 0.9955, 95% CI: 0.9969–0.9984, p < 0.0001). In simpler terms, as the level of i-tRF^GlyGCC increases in tissue samples, the level in serum samples also tends to increase proportionally. This strong correlation suggests that i-tRF^GlyGCC in serum might be a promising non-invasive biomarker for EOC (Fig. 2).

Fig. 2.Correlation of i-tRF^GlyGCC expression in tissues and serum of epithelial ovarian cancer (EOC) patients. Pearson’s correlation analysis demonstrates a strong positive correlation (r² = 0.9955, p < 0.0001) between i-tRF^GlyGCC expression levels in tumor tissues and serum samples. i-tRF^GlyGCC: internal tRNA-derived fragment Glycine-Glycine-Cytosine.

3.3 Prognostic significance of i-tRF^GlyGCC expression in epithelial ovarian cancer

In this study, we investigated the potential of i-tRF^GlyGCC as a prognostic biomarker for epithelial ovarian cancer (EOC) patients. Interestingly, we found a strong negative correlation between patient age and i-tRF^GlyGCC expression (r = −0.9911). In simpler terms, as patients’ age increased, their i-tRF^GlyGCC levels significantly decreased (p-value < 0.0001) (Fig. 3).

Fig. 3.Correlation between age and i-tRF^GlyGCC expression in epithelial ovarian cancer (EOC) tissues. Pearson’s correlation analysis reveals a strong positive correlation (r = 0.9824, p < 0.0001) between patient age and i-tRF^GlyGCC expression levels in EOC tissues. i-tRF^GlyGCC: internal tRNA-derived fragment Glycine-Glycine-Cytosine.

Our data indicates a strong correlation between i-tRF^GlyGCC expression and several prognostic features in EOC patients (Table 1).

Table 1.itRNA^GlyGCC expression in epithelial ovarian cancer according to different clinical and pathological features.

		No	itRNAGlyGCC	t-test	p
			Mean	SD
Stage
	IIIA	33	2.523	2.37	118.20*	<0.0001
	IIIB	30	3.613	0.94
	IIIC	83	8.236	2.18
Pathology
	Serous	91	4.078	2.28	4.97	<0.0001
	Non-serous	55	6.466	3.97
Grade
	Moderate + Well differentiated	59	2.602	1.14	19.30	<0.0001
	Poorly differentiated	87	8.296	2.06
Ascites
	Absent	57	2.525	1.09	18.87	<0.0001
	Present	89	8.217	2.10
Response
	CR, Microscopic disease, PR	75	3.197	1.54	21.59	<0.0001
	No response or progressive disease	71	8.951	1.68
Residual Disease
	>2 cm	57	8.217	2.10	18.87	<0.0001
	<2 cm	89	2.525	1.09
*ANOVA test. itRNAGlyGCC: internal tRNA-derived fragment Glycine-Glycine-Cytosine; SD: Stable Disease; CR: Complete Response; PR: Partial Response.

Stage: Patients with higher stages (IIIB & IIIC) have significantly higher i-tRF^GlyGCC levels compared to Stage IIIA (p-value < 0.0001). This suggests a potential link between the advanced disease stage and increased i-tRF^GlyGCC expression.

Pathology: Serous tumors, the most common type of EOC, have lower i-tRF^GlyGCC levels than non-serous tumors (p-value < 0.0001). This indicates a possible association between tumor type and i-tRF^GlyGCC expression.

Grade: Patients with poorly differentiated tumors (more aggressive) have significantly higher i-tRF^GlyGCC levels compared to those with moderately or well-differentiated tumors (p-value < 0.0001). This suggests a potential association between higher tumor grade and increased i-tRF^GlyGCC expression.

Ascites: Patients with ascites (fluid buildup in the abdomen) have significantly higher i-tRF^GlyGCC levels compared to those without ascites (p-value < 0.0001). This suggests a possible correlation between ascites and increased i-tRF^GlyGCC expression.

Response to Treatment: Patients with complete response (CR), microscopic disease, or partial response (PR) have significantly lower i-tRF^GlyGCC levels compared to those with no response or progressive disease (p-value < 0.0001). This suggests a potential link between better treatment response and lower i-tRF^GlyGCC expression.

Residual Disease: Patients with minimal residual disease (<2 cm) have significantly lower i-tRF^GlyGCC levels compared to those with larger residual tumors (>2 cm) (p-value < 0.0001). This suggests a possible association between lower residual disease burden and decreased i-tRF^GlyGCC expression.

These results highlight a promising link between i-tRF^GlyGCC expression and various prognostic features in EOC. Higher i-tRF^GlyGCC levels are associated with advanced stage, aggressive tumor characteristics and poorer treatment outcomes.

In the multiple regression analysis, we examined how the dependent variable A (EOC) relates to a set of independent variables. The model included ten independent variables: B (ANOT), C (Stage), D (Age), E (serum), F (Pathology), G (Grade), H (Ascites), I (Response) and J (Residual Disease). After examining the parameter estimates, we found that only three variables, C (Stage), F (Pathology) and I (Response), showed statistically significant relationships with the dependent variable. These variables had p-values below the threshold of 0.05, indicating a significant association (Table 2).

Table 2.Multiple regression analysis of different prognostic features.

Regression Statistics
Multiple R	1
R Square	1
Adjusted R Square	0.992701
Standard Error	1.65 × 10−15
Observations	146
ANOVA
	df	SS	MS	F	Significance F
Regression	9	1580.858	175.6509	7.28328 × 1031	0
Residual	137	3.72 × 10−28	2.71 × 10−30
Total	146	1580.858
	Coefficients	Standard Error	t Stat	p-value	Lower 95%	Upper 95%	Lower 95.0%	Upper 95.0%
Intercept	−6.9 × 10−15	1.05 × 10−15	−6.53821	1.13763 × 10−9	−8.9 × 10−15	−4.8 × 10−15	−8.9 × 10−15	−4.8 × 10−15
ANOT	−3 × 10−17	8.16 × 10−16	−0.0369	0.97061942	−1.6 × 10−15	1.58 × 10−15	−1.6 × 10−15	1.58 × 10−15
Stage	1.79 × 10−16	6.7 × 10−17	2.668292	0.008543895	4.63 × 10−17	3.11 × 10−16	4.63 × 10−17	3.11 × 10−16
Age	1	5.01 × 10−16	1.99 × 1015	0	1	1	1	1
Serum	−2.7 × 10−16	5.49 × 10−16	−0.49673	0.620173186	−1.4 × 10−15	8.13 × 10−16	−1.4 × 10−15	8.13 × 10−16
Pathology	2.23 × 10−16	4.91 × 10−17	4.554181	1.15064 × 10−5	1.26 × 10−16	3.2 × 10−16	1.26 × 10−16	3.2 × 10−16
Grade	2.87 × 10−17	1.23 × 10−16	0.232542	0.816464479	−2.2 × 10−16	2.73 × 10−16	−2.2 × 10−16	2.73 × 10−16
Ascites	2.02 × 10−17	1.13 × 10−16	0.178279	0.858767184	−2 × 10−16	2.44 × 10−16	−2 × 10−16	2.44 × 10−16
Response	1.36 × 10−16	5.4 × 10−17	2.509885	0.013240946	2.88 × 10−17	2.43 × 10−16	2.88 × 10−17	2.43 × 10−16
Residual Disease	0	0	65,535	0.817564432	0	0	0	0
ANOVA: Analysis of Variance; df: the degrees of freedom; SS: the sum of squares; MS: Mean Squared Errors; ANOT: Adjacent normal ovarian tissues.

3.4 i-tRF^GlyGCC expression and its association with survival outcomes in EOC patients

High expression of i-tRF^GlyGCC is linked to poorer overall and disease-free survival in patients with EOC. Patients with high i-tRF^GlyGCC expression had a median overall survival of 36 months, while those with low expression had 61 months (p = 0.0032), indicating a 25-month difference in median survival time. Likewise, the median disease-free survival was 42 months for high-expression patients and 61 months for low-expression patients (p = 0.0128), suggesting that patients with high i-tRF^GlyGCC expression experienced disease recurrence on average 19 months earlier (Fig. 4).

These findings suggest that i-tRF^GlyGCC expression could serve as a prognostic marker for EOC survival.

Fig. 4.Survival analysis based on i-tRF^GlyGCC expression in epithelial ovarian cancer (EOC) patients. (A) Overall survival (OS) in EOC patients stratified by high and low i-tRF^GlyGCC expression levels (p = 0.0032). (B) Disease-free survival (DFS) in EOC patients stratified by high and low i-tRF^GlyGCC expression levels (p = 0.0128). High i-tRF^GlyGCC expression is associated with significantly poorer survival outcomes. i-tRF^GlyGCC: internal tRNA-derived fragment Glycine-Glycine-Cytosine.

4. Discussion

Ovarian cancer is a highly lethal malignancy, primarily due to the challenges associated with its early diagnosis [24]. In its initial stages, the disease often presents with nonspecific symptoms, such as bloating, pelvic pain and changes in bowel habits, which are easily mistaken for benign conditions [25]. This lack of distinct clinical manifestations frequently results in delayed or misdiagnosis, significantly worsening patient prognosis [26]. Furthermore, the absence of reliable and accessible screening methods exacerbates the difficulty of early detection. Unlike breast or cervical cancer, for which routine screening tests are well-established, ovarian cancer lacks widely available diagnostic tools, allowing the disease to progress undetected until advanced stages [26]. This underscores the critical need for the development of effective early detection strategies to improve outcomes for patients with ovarian cancer.

Recent research has highlighted the potential of i-tRF^GlyGCC, a transfer RNA (tRNA)-derived fragment, as a promising biomarker in various cancers. Studies have shown significantly elevated levels of i-tRF^GlyGCC in the blood plasma of colorectal cancer (CRC) patients compared to healthy individuals, suggesting its potential utility in developing non-invasive blood-based diagnostic tests for CRC [27]. Additionally, research has identified a positive correlation between increased i-tRF^GlyGCC expression and poorer overall and disease-free survival rates in patients with chronic lymphocytic leukemia (CLL), underscoring its prognostic value [28]. These findings indicate that i-tRF^GlyGCC could serve as a valuable tool for early cancer detection, risk stratification, and the development of personalized treatment strategies, offering new avenues for improving cancer management and patient outcomes.

In this study, we investigated the expression levels of i-tRF^GlyGCC in both tumor tissues and serum samples from epithelial ovarian cancer (EOC) patients to evaluate its potential as a diagnostic and prognostic biomarker.

The expression of i-tRF^GlyGCC was significantly downregulated in epithelial ovarian cancer (EOC) tissues compared to normal ovarian tissues, with a statistically significant difference (p < 0.0001). Consistent findings were observed in serum samples, where i-tRF^GlyGCC levels were significantly lower in ovarian cancer patients compared to healthy controls, further demonstrating a statistically significant difference (p < 0.0001).

Our study explored the relationship between i-tRF^GlyGCC levels in epithelial ovarian cancer (EOC) tissues and corresponding serum samples. Notably, we identified a strong positive correlation between i-tRF^GlyGCC expression in tissues and serum, indicating that elevated levels in tissue samples were associated with similarly increased levels in the blood. This close correlation suggests that measuring i-tRF^GlyGCC in serum could serve as a reliable, non-invasive method for diagnosing EOC. These findings underscore the potential of serum i-tRF^GlyGCC as a promising non-invasive biomarker for epithelial ovarian cancer, offering a practical alternative to tissue biopsies, which are more invasive and challenging to obtain.

A study by Christodoulou et al. [7] investigated the prognostic significance of the tRNA fragment i-tRF^GlyGCC in colorectal cancer (CRC) patients. By analyzing total RNA from 91 cancerous tissue samples and 83 adjacent normal tissues, the study revealed significantly reduced levels of i-tRF^GlyGCC in CRC tissues compared to normal colorectal tissues. Similarly, another study explored the potential of plasma exosomal tRNA-derived fragments (tRFs) as diagnostic biomarkers for non-small cell lung cancer (NSCLC). The study identified a specific panel of tRFs, including tRF-Leu-TAA-005, tRF-Asn-GTT-010, tRF-Ala-AGC-036, tRF-Lys-CTT-049 and tRF-Trp-CCA-057, which were significantly downregulated in NSCLC patients compared to healthy individuals [29]. Additionally, research on bladder cancer examined tRFs in urine and blood extracellular vesicles (EVs) as noninvasive biomarkers. Distinct tRF profiles were observed in urine EVs from cancer patients compared to healthy individuals, and serum tRF levels changed following surgery, highlighting their potential for bladder cancer diagnosis and monitoring [30]. Furthermore, Li et al. [31] studied tRF-Pro-CGG in pancreatic ductal adenocarcinoma (PDAC) and found significantly lower levels of tRF-Pro-CGG in PDAC patients, underscoring its potential role in this malignancy. Collectively, these studies demonstrate the growing interest in tRFs as promising diagnostic and prognostic biomarkers across various cancer types.

The current study identified a strong correlation between i-tRF^GlyGCC expression and several indicators of poor prognosis in epithelial ovarian cancer (EOC). Patients with advanced-stage disease, non-serous histology, higher tumor grades, ascites, poorer treatment response, and larger residual tumors exhibited significantly elevated levels of i-tRF^GlyGCC (p < 0.0001). These findings suggest that i-tRF^GlyGCC may serve as a potential biomarker for aggressive EOC and unfavorable clinical outcomes. Similarly, in pancreatic ductal adenocarcinoma (PDAC), Li et al. [31] reported that lower levels of tRF-Pro-CGG were associated with poorer prognosis and reduced survival, highlighting its potential as a biomarker for disease progression and treatment guidance. In gastric cancer, Gan et al. [32] demonstrated that dysregulation of tsRNAs was closely linked to adverse clinicopathological factors, including lymph node metastasis, advanced Tumor-Node-Metastasis (TNM) stage, larger tumor size and vascular invasion. Additionally, Zhang et al. [33] identified three specific tRNA fragments (tRFs)—tRF-Gly-CCC-046, tRF-Tyr-GTA-010 and tRF-Pro-TGG-001—that were significantly downregulated in both tissue and blood samples from breast cancer patients compared to healthy individuals, suggesting their potential as biomarkers for early breast cancer detection. Collectively, these studies underscore the emerging role of tRNA-derived fragments as valuable biomarkers for cancer diagnosis, prognosis, and therapeutic decision-making across various malignancies.

In our study, we observed that epithelial ovarian cancer (EOC) patients with elevated levels of i-tRF^GlyGCC faced a significantly worse prognosis. Specifically, these patients had a median overall survival (OS) that was 25 months shorter (36 months vs. 61 months) and experienced disease recurrence 19 months earlier compared to those with lower i-tRF^GlyGCC levels. Similarly, in colorectal cancer (CRC), patients with high i-tRF^GlyGCC levels exhibited significantly poorer outcomes, including shorter disease-free intervals (DFS) and overall survival (OS), compared to those with lower levels [7]. Consistent with these findings, Karousi et al. [18] reported that increased levels of i-tRF^GlyGCC were associated with poor overall survival in chronic lymphocytic leukemia (CLL) patients. These results collectively highlight the prognostic significance of i-tRF^GlyGCC across multiple cancer types, underscoring its potential as a biomarker for predicting aggressive disease and unfavorable clinical outcomes.

This study identifies i-tRF^GlyGCC as a promising biomarker for the diagnosis and prognosis of epithelial ovarian cancer (EOC). However, further research is required to validate and confirm these findings, ensuring their reproducibility and clinical applicability.

5. Limitations of the study

Our study has several limitations, including a relatively small and homogeneous sample size of 142 stage III epithelial ovarian cancer patients, which may not fully represent the broader population or earlier disease stages. Conducted at a single institution (Zagazig University Hospitals), the findings may lack generalizability due to potential institutional biases and limited geographic or ethnic diversity. The study also lacked long-term follow-up data, which could provide deeper insights into the prognostic value of i-tRF^GlyGCC over time. Additionally, potential confounding factors such as comorbidities, lifestyle, and genetic predispositions were not accounted for, and the reliance on qPCR for measuring i-tRF^GlyGCC levels may introduce technical variability. As an exploratory study, the findings require validation in larger, multicenter cohorts, and the absence of functional studies limits the understanding of the biological mechanisms underlying i-tRF^GlyGCC’s role in ovarian cancer progression. These limitations underscore the need for further research to confirm and expand upon these results.

6. Conclusions

In conclusion, this study presents compelling evidence for i-tRF^GlyGCC as a promising novel biomarker in epithelial ovarian cancer. Our findings demonstrate significantly reduced i-tRF^GlyGCC expression in both tumor tissues and serum of EOC patients compared to healthy controls, with perfect diagnostic accuracy as indicated by ROC curve analysis. The strong associations between i-tRF^GlyGCC expression and adverse clinicopathological features, including advanced stage, poor differentiation and presence of ascites, alongside its correlation with diminished survival outcomes, suggest its potential value as both a diagnostic and prognostic marker. These results provide a strong foundation for the clinical application of i-tRF^GlyGCC in EOC management, though larger multicenter studies are needed to validate these findings and establish standardized cutoff values for clinical implementation. Our study contributes to the growing understanding of tRNA-derived fragments in cancer biology and opens new avenues for improving EOC patient care through enhanced early detection and more accurate prognostic assessment.

Availability of data and materials

All data will be available upon request.

Author contributions

HD, AFG, WHE—Conceptualization, Methodology, Software. AAA, AA, MMB, RAK—Data curation, Writing-Original draft preparation. AAA, AFG, WHE—Supervision. AAA—Software, Validation. TMA, AFG, WHE—Writing-Reviewing and Editing. All authors have read the manuscript and approved the submission.

Ethics approval and consent to participate

All participants provided written informed consent after Zagazig University’s Research Ethics Committee granted authorization (ZU-232/2018).

Acknowledgment

The authors extend their appreciation to Taif University, Saudi Arabia, for supporting this work through project No. TU-DSPP-2024-54.

Funding

This research was funded by Taif University, Saudi Arabia, project No. TU-DSPP-2024-54.

Conflict of interest

The authors declare no conflict of interest.