Hereditas

Activation of TRIM37 by ATF6 and degradation of ACSL4: inhibiting ferroptosis and propelling cervical cancer progression

Abstract Background Cervical cancer (CC), a prevalent gynecological malignancy, shows high global incidence and mortality. Tripartite motif-containing 37 (TRIM37), a significant ubiquitinating enzyme, is overexpressed in CC, fueling its progression, but its role in ferroptosis here is unknown. Methods TRIM37 expression in CC tissues was first predicted using bioinformatics software. Then, RT-qPCR and Western blot were utilized to confirm TRIM37 expression in CC tissues and cells. Subsequently, cellular behaviors were examined by EdU, flow cytometry, and Transwell assay. Besides, ferroptosis-related indicators were detected by using corresponding kits. The dual luciferase reporter assay was conducted to identify the binding between TRIM37 and Activating Transcription Factor 6 (ATF6). Additionally, the Co-IP assay was applied to validate the interaction between TRIM37 and Acyl-CoA Synthetase Long-Chain Family Member 4 (ACSL4). Finally, the functions of TRIM37 in vivo were investigated by establishing a xenograft tumor model. Results TRIM37 expression was increased in CC tissues and cells. Silencing TRIM37 suppressed cell malignant behaviors and promoted ferroptosis. ATF6 activated TRIM37 transcription, with TRIM37 upregulation counteracting ATF6 knockdown effects. TRIM37 degraded ACSL4, and silencing ACSL4 reversed TRIM37 knockdown effects. TRIM37 overexpression counteracted ATF6 knockdown’s impact on tumor growth in vivo. Conclusion ATF6 regulated the expression of TRIM37, which in turn promoted the ubiquitination and degradation of ACSL4, facilitating the progression of CC. Graphical Abstract

A novel chromatin regulator-related immune checkpoint related gene prognostic signature and potential candidate drugs for endometrial cancer patients

Abstract Background Endometrial cancer (EC) is the most common gynecologic malignancy in developed countries and its prevalence is increasing. As an emerging therapy with a promising efficacy, immunotherapy has been extensively applied in the treatment of solid tumors. In addition, chromatin regulators (CRs), as essential upstream regulators of epigenetics, play a significant role in tumorigenesis and cancer development. Methods CRs and immune checkpoint-related genes (ICRGs) were obtained from the previous top research. The Genome Cancer Atlas (TCGA) was utilized to acquire the mRNA expression and clinical information of patients with EC. Correlation analysis was utilized for screen CRs-related ICRGs (CRRICRGs). By Cox regression and least absolute shrinkage and selection operator (LASSO) analysis, prognosis related CRRICRGs were screened out and risk model was constructed. The Kaplan–Meier curve was used to estimate the prognosis between high- and low-risk group. By comparing the IC50 value, the drugs sensitivity difference was explored. We obtained small molecule drugs for the treatment of UCEC patients based on CAMP dataset. Results We successfully constructed a 9 CRRICRs-based prognostic signature for patients with UCEC and found the riskscore was an independent prognostic factor. The results of functional analysis suggested that CRRICRGs may be involved in immune processes associated with cancer. Immune characteristics analysis provided further evidence that the CRRICRGs-based model was correlated with immune cells infiltration and immune checkpoint. Eight small molecule drugs that may be effective for the treatment of UCEC patients were screened. Effective drugs identified by drug sensitivity profiling in high- and low-risk groups. Conclusion In summary, our study provided novel insights into the function of CRRICRGs in UCEC. We also developed a reliable prognostic panel for the survival of patients with UCEC.

circ-NOLC1 inhibits the development of cervical cancer by regulating miR-330-5p-PALM signaling axis

Abstract Background Recent studies have increasingly demonstrated that circular RNAs (circRNAs) play significant roles in the occurrence and progression of cervical cancer (CC). In CC, circRNAs act as ceRNAs by sponging miRNAs to regulate genes associated with proliferation, migration, and apoptosis, exhibiting both promoting and inhibiting effects on tumor progression. The aim of this study was to clarify the role of hsa_circ_0019686 (named circ-NOLC1) in CC. Methods By conducting an online GEO2R analysis of the expression profile GSE113696 in the GEO database, circ-NOLC1 was selected. The expression levels of circ-NOLC1 in CC cell lines were measured using real-time quantitative PCR (RT-qPCR). The role of circ-NOLC1 in CC was validated through both in vitro and in vivo gain-of-function assays. Bioinformatic analysis, combined with luciferase reporter and RNA Immunoprecipitation (RIP) assays, confirmed that circ-NOLC1 acts as a sponge for miR-330-5p and regulates the expression of paralemmin-1 (PALM). The role of the circ-NOLC1-miR-330-5p-PALM signaling axis in CC was elucidated through the rescue experiments. Relative gene expression levels were measured using RT-qPCR, while relative protein levels were assessed through immunohistochemistry (IHC). CCK-8, wound healing, Transwell, and flow cytometry assays were employed to evaluate CC cell proliferation, migration, and invasion, respectively. Results The expression levels of circ-NOLC1 were dramatically downregulated in CC cells (P < 0.001). Up-regulation of circ-NOLC1 significantly inhibited cell proliferation (P < 0.001), migration (P < 0.01) and invasion (P < 0.01), while promoting cell apoptosis (P < 0.001). In vivo studies showed that up-regulation of circ-NOLC1 suppressed tumor growth (tumor volume: P < 0.001; tumor weight: P < 0.01). Additionally, miR-330-5p was found to be up-regulated in CC (P < 0.001), whereas PALM was downregulated in CC (P < 0.001). The up-regulation of circ-NOLC1 inhibited the expression of miR-330-5p (P < 0.001) and enhanced the expression of PALM (P < 0.001). Rescue experiments further demonstrated that the up-regulation of circ-NOLC1 inhibited CC cell proliferation (P < 0.001), migration (P < 0.001), invasion (P < 0.001), while promoting apoptosis (P < 0.001) through the regulation of the miR-330-5p-PALM pathway. Conclusion The circ-NOLC1 inhibits CC development through regulating the miR-330-5p-PALM signaling axis. This finding reveals a novel mechanism and identifies potential therapeutic targets, emphasizing the necessity for further regulatory studies and clinical validation.

Identification of N6-methyladenosine-associated ferroptosis biomarkers in cervical cancer

Abstract Background Cervical cancer (CC) stands as a major contributor to female mortality. The pathogenesis of CC is linked with various factors. Our research aimed to unravel the underlying mechanisms of ferroptosis and m6A RNA methylation in CC through bioinformatics analysis. Methods Three CC datasets, including GSE9750, GSE63514, and TCGA-CESC, were incorporated. m6A-related genes were derived from published sources, while ferroptosis-related genes were obtained from the FerrDb database. Differential expression and correlation analyses were performed to identify differentially expressed m6A-related ferroptosis genes (DE-MRFGs) in CC. Subsequently, the biomarkers were further identified using machine learning techniques. Gene Set Enrichment Analysis (GSEA) and Kaplan–Meier (KM) survival analysis were also performed to comprehend these biomarkers. Furthermore, a competing endogenous RNAs (ceRNA) network involving biomarkers was established. Finally, biomarkers expression were verified by real-time quantitative polymerase chain reaction (RT-qPCR). Results From the DE-MRFGs, six genes, including ALOX12, EZH2, CA9, CDCA3, CDC25A, HSPB1, were selected. A nomogram constructed based on these biomarkers exhibited potential clinical diagnostic value for CC, with good diagnostic accuracy confirmed through calibration curves. GSEA unveiled associations of these biomarkers with cell proliferation, spliceosome, and base excision repair. KM survival analysis demonstrated significant differences in survival outcomes between high and low expressions of HSPB1, EZH2, and CA9 samples. A ceRNA network was constructed involving three biomarkers, such as CDC25A, CDCA3, and EZH2, 29 miRNAs, and 25 lncRNAs. In RT-qPCR verification, the expression of ALOX12, EZH2 and CDC25A was significantly higher in CC samples, while HSPB1 expression was higher in control samples. Conclusion Six genes, namely ALOX12, EZH2, CA9, CDCA3, CDC25A, and HSPB1, were identified as m6A-regulated ferroptosis biomarkers in CC. These findings offer valuable insights into disease pathogenesis and hold promise for advancing CC treatment and prognosis.

Development of a risk model based on autophagy-related genes to predict survival and immunotherapy response in ovarian cancer

Abstract Background Autophagy is a highly conserved cellular proteolytic process that can interact with innate immune signaling pathways to affect the growth of tumor cells. However, the regulatory mechanism of autophagy in the tumor microenvironment, drug sensitivity, and immunotherapy is still unclear. Methods Based on the prognostic autophagy-related genes, we used the unsupervised clustering method to divide 866 ovarian cancer samples into two regulatory patterns. According to the phenotypic regulation pattern formed by the differential gene between the two regulation patterns, a risk model was constructed to quantify patients with ovarian cancer. Then, we systematically analyzed the relationship between the risk model and immune cell infiltration, immunotherapeutic response, and drug sensitivity. Results Based on autophagy-related genes, we found two autophagy regulation patterns, and confirmed that there were differences in prognosis and immune cell infiltration between them. Subsequently, we constructed a risk model, which was divided into a high-risk group and a low-risk group. We found that the high-risk group had a worse prognosis, and the main infiltrating immune cells were adaptive immune cells, such as Th2 cells, Tgd cells, eosinophils cells, and lymph vessels cells. The low-risk group had a better prognosis, and the most infiltrated immune cells were innate immune cells, such as aDC cells, NK CD56dim cells, and NK CD56bright cells. Furthermore, we found that the risk model could predict chemosensitivity and immunotherapy response, suggesting that the risk model may help to formulate personalized treatment plans for patients. Conclusions Our study comprehensively analyzed the prognostic potential of autophagy-related risk models in ovarian cancer and determined their clinical guiding role in targeted therapy and immunotherapy.

N6-methyladenosine-related lncRNAs is a potential marker for predicting prognosis and immunotherapy in ovarian cancer

Abstract Background With a lack of specific symptoms, ovarian cancer (OV) is often diagnosed at an advanced stage. This coupled with inadequate prognostic indicators and treatments with limited therapeutic effect make OV the deadliest type of gynecological tumor. Recent research indicates that N6-methyladenosine (m6A) and long-chain non-coding RNA (lncRNA) play important roles in the prognosis of OV and the efficacy of immunotherapy. Results Using the Cancer Genome Atlas (TCGA) OV-related data set and the expression profiles of 21 m6A-related genes, we identified two m6A subtypes, and the differentially expressed genes between the two. Based on the differentially expressed lncRNAs in the two m6A subtypes and the lncRNAs co-expressed with the 21 m6A-related genes, single-factor cox and LASSO regression were used to further isolate the 13 major lncRNAs. Finally, multi-factor cox regression was used to construct a m6A-related lncRNA risk score model for OV, with good performance in patient prognosis. Using risk score, OV tumor samples are divided into with high- and low-score groups. We explored the differences in clinical characteristics, tumor mutational burden, and tumor immune cell infiltration between the two groups, and evaluated the risk score’s ability to predict the benefit of immunotherapy. Conclusion Our m6A-based lncRNA risk model could be used to predict the prognosis and immunotherapy response of future OV patients.

COL1A1, ITGB1, THY1, and PDGFRA: key immune-related genes in uterine corpus endometrial carcinoma with prognostic and therapeutic implications

Abstract Uterine corpus endometrial carcinoma (UCEC) is one of the most common gynecological malignancies, characterized by complex molecular alterations that drive its progression. Understanding the molecular mechanisms underlying UCEC is crucial for developing effective diagnostic, prognostic, and therapeutic strategies. Immune-related genes, such as COL1A1, ITGB1, THY1, and PDGFRA, have been implicated in various cancers, but their roles in UCEC remain underexplored. In this study, we investigate the roles of these genes in the development and progression of UCEC. Using both in silico and in vitro approaches, we found that these genes were dysregulated in UCEC. Our results revealed the downregulation of COL1A1, ITGB1, THY1, and PDGFRA in UCEC compared to normal tissues. Further, promoter methylation analysis showed increased methylation of these genes in UCEC. Survival analysis highlighted their potential as prognostic markers, with lower expression linked to poor patient survival. Additionally, genetic alteration analysis demonstrated mutations in these genes across UCEC patients. Our results also showed that overexpression of COL1A1 in KLE and HEC-1B cells significantly reduced cell proliferation, colony formation, and migration, indicating that COL1A1 overexpression impacts critical cellular behaviors in UCEC. Finally, we explored the therapeutic potential of targeting these genes, suggesting that they may offer valuable insights for personalized treatment strategies in UCEC. This study identifies COL1A1, ITGB1, THY1, and PDGFRA as crucial regulators of UCEC progression, with altered expression linked to tumor behavior and patient survival. Overexpression of COL1A1 impaired cell proliferation, colony formation, and migration. Future research should focus on elucidating the molecular mechanisms of these genes, exploring their therapeutic targeting in preclinical models, and validating their clinical potential as biomarkers in larger patient cohorts to improve treatment strategies for UCEC.

Key wound healing genes as diagnostic biomarkers and therapeutic targets in uterine corpus endometrial carcinoma: an integrated in silico and in vitro study

Abstract Background Uterine Corpus Endometrial Carcinoma (UCEC) is a prevalent gynecologic malignancy with complex molecular underpinnings. This study identifies key woundhealing genes involved in UCEC and elucidates their roles through a comprehensive analysis. Methods In silico and in vitro experiments. Results Seventy wound healing-associated genes were extracted from the Gene Ontology (GO) database, and a protein-protein interaction (PPI) network was constructed using the STRING database. CytoHubba analysis in Cytoscape identified six pivotal hub genes: CD44, FGF2, FGF10, KDM6A, FN1, and MMP2. These genes exhibited significantly lower expression in UCEC cell lines compared to normal controls, as confirmed by RT-qPCR. Receiver Operating Characteristic (ROC) analysis demonstrated their potential as diagnostic biomarkers, with Area Under the Curve (AUC) values ranging from 0.94 to 1.00. Validation using TCGA datasets revealed consistent downregulation of these genes in UCEC samples, corroborated by immunohistochemical staining. Promoter methylation analysis showed significantly higher methylation levels in UCEC, correlating with decreased mRNA expression and poor survival outcomes. Genetic alteration analysis indicated frequent mutations in FN1 and KDM6A, although these did not significantly affect survival. Functional analysis using the CancerSEA database highlighted the involvement of these genes in critical cancer-related processes, including angiogenesis, apoptosis, and metastasis. Immune correlation studies revealed significant associations with immune inhibitor genes and distinct expression patterns across immune subtypes. Overexpression studies in UCEC cell lines demonstrated that CD44 and MMP2 reduce proliferative ability while enhancing migration and wound healing. Conclusion Collectively, these findings underscore the crucial roles of CD44, FGF2, FGF10, KDM6A, FN1, and MMP2 in UCEC pathogenesis, highlighting their potential as biomarkers and therapeutic targets in this malignancy.

METTL3-dependent DLG2 inhibits the malignant progression of cervical cancer by inactivating the Hippo/YAP signaling

Abstract Background Discs large homolog 2 (DLG2) has been implicated in cancer development, yet its role in cervical cancer remains unclear. This study aims to explore the regulatory mechanism of DLG2 in cervical cancer and its clinical implications. Methods Quantitative reverse transcription polymerase chain reaction and western blotting assays were employed to detect RNA and protein expression, respectively. Colony formation assay, 5-Ethynyl-2’-deoxyuridine assay, flow cytometry, and transwell assays were conducted for cell functional analysis. A xenograft mouse model assay was performed to analyze tumor tumorigenesis in vivo. m6A RNA immunoprecipitation assay was used to analyze the association of METTL3 and DLG2. Results DLG2 was underexpressed in cervical cancer tissues and cells. Elevating DLG2 levels significantly suppressed cervical cancer cell proliferation, migration, and invasion, while promoting apoptosis. Additionally, DLG2 overexpression led to the deactivation of the Hippo/YAP signaling pathway. In vivo, DLG2 overexpression was shown to reduce tumor formation. We also discovered that METTL3 destabilized DLG2 mRNA through an m6A-dependent mechanism. Moreover, lowering DLG2 expression mitigated the effects of METTL3 silencing on cervical cancer cell malignancy. Conclusion DLG2 acted as a tumor suppressor in cervical cancer by inhibiting the Hippo/YAP signaling pathway. The METTL3-dependent regulation of DLG2 mRNA stability could be a critical factor in cervical cancer progression.

METTL3 mediates m6A methylation modification of ULBP2 and affects the progression of cervical cancer

Abstract Background Cervical cancer (CC) is one of the most prevalent malignancies in women, posing a significant challenge globally. However, the precise molecular mechanism regulating CC progression through methyltransferase-like protein 3 (METTL3) and UL16 Binding Protein 2 (ULBP2) remains largely unknown. Methods Bioinformatic analysis was used to identify the effect of ULBP2 expression in CC tissues. RT-qPCR and western blotting were employed to assess the mRNA and protein expression in CC cells and tissues. Methylthiazolyldiphenyl-tetrazolium bromide (MTT), 5‑Ethynyl‑2’‑deoxyuridine (EdU), wound healing, and transwell assays were utilized to estimate cell viability, proliferation, and metastasis, respectively. Cell apoptosis was detected by flow cytometry. CC cells were treated with different doses of radiotherapy. The m6A level was measured using methylated RNA immunoprecipitation (MeRIP) assay. A xenograft assay was conducted to further verify the roles of ULBP2 in CC. Results ULBP2 was upregulated in CC. Downregulation of ULBP2 restrained the proliferation, metastasis and radiotherapy resistance of CC cells. METTL3 regulated m6A methylation modification of ULBP2. Insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1) promoted m6A methylation modification of ULBP2. METTL3 influenced the expression of ULBP2 and impacted the biological function of the CC cells. Silencing ULBP2 reduced the radioresistance of CC in vivo. Radiotherapy altered the gut microbiota in CC patients. Conclusion METTL3 modulated the m6A methylation of ULBP2, affecting the oncogenic properties and radioresistance of CC cells.

Prognostic value of Linc00662/miR-16-5p/FASN in cervical cancer and regulation of tumor progression

Abstract Background Cervical cancer (CC) is the world's single most frequent gynecological cancer, is more than 500,000 new annual cases globally, and is a serious threat to women's reproductive health. LncRNAs have significant effects on human diseases; nevertheless, the expression of Linc00662 in CC and its mechanism of action are not yet entirely clear. The goal of the work was to investigate the expression, prognostic value and biological utility of Linc00662 in CC progression and to identify its underlying mechanisms in molecular terms. Methods Expression levels of Linc00662, miR-16-5p and FASN in CC tissues and cells were detected through real-time quantitative PCR. Determination of cell proliferative capacity by CCK-8. Cell migration and invasion were assessed through Transwell assay. Binding of Linc00662 to miR-16-5p was mediated through a dual-luciferase reporter gene test was validated. Results Linc00662 expression levels were significantly elevated in CC. High Linc00662 expression was strongly linked to increased tumor size, later FIGO staging, poorer tumor differentiation, mesenchymal infiltration, and lymph node metastasis, and high Linc00662 expression predicted a poor prognosis. Silencing Linc00662 reduced the proliferation, migration, and invasion of CC cells. Furthermore, Linc00662 negatively regulated miR-16-5p and indirectly regulated the upregulation of FASN expression. Conclusions Linc00662 positively regulates FASN expression through targeting miR-16-5p and facilitates CC cell proliferation, migration and invasion, promoting CC progression.

Mir-615-5p inhibits cervical cancer progression by targeting TMIGD2

Abstract Background Cervical cancer (CC) is a prevalent gynecological malignancy, contributing to a substantial number of fatalities among women. MicroRNAs (miRNAs) have emerged as promising biomarkers with significant potential for the early detection and prognosis of CC. Objective This study aimed to explore the clinical significance and biological role of miR-615-5p in CC, with the goal of identifying novel biomarkers for this disease. Materials and methods The levels of miR-615-5p and TMIGD2 mRNA in tissue samples and cells were quantified through quantitative reverse transcription real-time PCR, followed by statistical analyses to investigate the correlation between miR-615-5p and clinical data. The effects of miR-615-5p on the proliferation and metastasis of CC cells were evaluated using the Cell Counting Kit-8 and Transwell assays. The potential mechanism of miR-615-5p was elucidated by bioinformatics analyses and Dual-luciferase reporter assay. Western blotting was employed to measure the protein levels of TMIGD2. Results In CC, the downregulation of miR-615-5p was related to poor prognosis and emerged as an independent prognostic factor. The levels of miR-615-5p were reduced in CC cells. miR-615-5p overexpression restrained the proliferation and metastasis of CC cells. Furthermore, TMIGD2 was identified as a target gene regulated by miR-615-5p, and its expression was notably elevated in CC. The influence of miR-615-5p on the biological behaviors of CC cells was mediated through the modulation of TMIGD2. Conclusions Downregulation of miR-615-5p was a prognostic indicator of poor prognosis in CC. miR-615-5p exerted its tumor-suppressive effects by inhibiting cell growth and metastasis through the regulation of TMIGD2.

Integrated analysis of N6-methyladenosine- and 5-methylcytosine-related long non-coding RNAs for predicting prognosis in cervical cancer

Abstract Background N6-methyladenosine (m6A) and 5-methylcytosine (m5C) play a role in modifying long non-coding RNAs (lncRNAs) implicated in tumorigenesis and progression. This study was performed to evaluate prognostic value of m6A- and m5C-related lncRNAs and develop an efficient model for prognosis prediction in cervical cancer (CC). Methods Using gene expression data of TCGA set, we identified m6A- and m5C-related lncRNAs. Consensus Clustering Analysis was performed for samples subtyping based on survival-related lncRNAs, followed by analyzing tumor infiltrating immune cells (TIICs). Optimal signature lncRNAs were obtained using lasso Cox regression analysis for constructing a prognostic model and a nomogram to predict prognosis. Results We built a co-expression network of 23 m6A-related genes, 15 m5C-related genes, and 62 lncRNAs. Based on 9 m6A- and m5C-related lncRNAs significantly associated with overall survival (OS) time, two molecular subtypes were obtained, which had significantly different OS time and fractions of TIICs. A prognostic model based on six m6A- and m5C-related signature lncRNAs was constructed, which could dichotomize patients into two risk subgroups with significantly different OS time. Prognostic power of the model was successfully validated in an independent dataset. We subsequently constructed a nomogram which could accurately predict survival probabilities. Drug sensitivity analysis found preferred chemotherapeutic agents for high and low-risk patients, respectively. Conclusion Our study reveals that m6A- and m5C-related lncRNAs are associated with prognosis and immune microenvironment of CC. The m6A- and m5C-related six-lncRNA signature may be a useful tool for survival stratification in CC and open new avenues for individualized therapies.

miR-423-5p mediates LINC00886 regulation of ovarian cancer aggressiveness and immune evasion via the TLR4/Myd88/NF-κB/PD-L1 pathway

Abstract Background Ovarian cancer has poor treatment outcomes. This study aims to explore the clinical importance of LINC00886 and its effects on cancer cell behavior in ovarian cancer, potentially offering a new therapeutic target. Materials and methods RT-qPCR was used to detect LINC00886 expression in ovarian cancer tissue, with analysis of clinicopathological data and prognosis based on LINC00886 expression levels. CCK-8, Traswell, and Annexin V-FITC/PI flow cytometry assays were used to evaluate the impact of molecular expression on cell viability, invasiveness, and apoptosis. RIP and dual luciferase reporter gene assays were used to validate interactions among miR-423-5p, LINC00886, and TLR4. Western blot analysis was conducted to investigate downstream signaling proteins, and ELISA was used to measure TNF-α and IFN-γ levels in cell co-culture. Results LINC00886 is upregulated in ovarian cancer tissues and cell lines, and its high expression is associated with poor prognosis; downregulating LINC00886 inhibits cell viability and invasiveness while inducing apoptosis. miR-423-5p is downstream of LINC00886 and upstream of TLR4. Inhibiting miR-423-5p reverses the suppressive effects of LINC00886 downregulation on cancer cell behavior. Overexpressing TLR4 enhances cellular processes. Furthermore, downregulating LINC00886 reduces the expression of TLR4, Myd88, phosphorylated NF-κB p65, and PD-L1, while increasing TNF-α and IFN-γ levels and enhancing CD8 + T cell antitumor activity, thereby reducing tumor cell immune escape. Conclusions LINC00886 drives ovarian cancer progression and immune escape through themiR-423-5p/TLR4/Myd88/NF-κB/PD-L1 axis, establishing its potential as both a prognostic biomarker and therapeutic target.

Multi-Omics analysis and in vitro validation reveal diagnostic and therapeutic roles of novel hub genes in ovarian cancer

Abstract Ovarian cancer (OC) remains a highly lethal gynecologic malignancy due to late diagnosis and limited therapeutic options. In this study, we aimed to identify and functionally validate novel hub genes associated with OC progression. We integrated four GEO microarray datasets (GSE54388, GSE40595, GSE18521, and GSE12470) to identify differentially expressed genes (DEGs) between OC and healthy tissues using the limma package. A total of 22 common DEGs were identified, of which four—SNRPA1, LSM4, TMED10, and PROM2—emerged as hub genes based on PPI network centrality. Expression analyses using TCGA data and RT-qPCR confirmed the significant upregulation of these genes in OC samples. Promoter methylation analysis showed hypomethylation in tumors, while ROC analysis revealed high diagnostic accuracy (AUC = 1.0). Although these genes were not significantly associated with overall survival in meta-analysis, they were strongly involved in oncogenic pathways such as EMT, apoptosis, and DNA repair. Predicted miRNAs (e.g., hsa-miR-1178-5p and hsa-miR-31-5p) targeting hub genes were significantly downregulated in OC cell lines. Immune analysis indicated that hub gene expression was correlated with immune subtypes, checkpoint inhibitors, and reduced immune infiltration. Drug sensitivity analysis suggested that high expression of TMED10 and PROM2 may confer susceptibility to chemotherapeutic agents. Functional assays following siRNA-mediated knockdown of TMED10 and PROM2 in A2780 and OVCAR3 cells revealed significant reductions in proliferation, colony formation, and migration. These findings highlight SNRPA1, LSM4, TMED10, and PROM2 as potential diagnostic markers and therapeutic targets in OC, warranting further investigation for clinical translation.

LncRNA LUCAT1 as a prognostic biomarker in cholangiocarcinoma through targeting miR-141-3p: clinical and functional insights

Abstract Background Cholangiocarcinoma (CHOL) has a poor prognosis due to its asymptomatic progression, challenges in early detection, and limited treatment options. The lncRNA LUCAT1 is highly expressed in several cancers, including lung, gastric, ovarian, and osteosarcoma tissues. Aim This study investigates the potential of LUCAT1 as a diagnostic and prognostic biomarker for CHOL. Materials and methods In this study, we collected tumor tissues and adjacent tumor healthy tissues from 83 CHOL patients. LUCAT1 expression was quantified in CHOL tissues and cell lines via RT-qPCR. Diagnostic and prognostic significance was assessed through ROC curves, Kaplan-Meier survival analysis, and Cox regression models. The biological effects of LUCAT1 on cell proliferation and migration were examined using QBC939 and HuCCT1 cells with transfection assays. The regulatory interaction between LUCAT1 and miR-141-3p was validated using a dual-luciferase reporter assay. Results Elevated expression of LUCAT1 was observed in CHOL tumor tissues and human cholangiocarcinoma cells, correlating with tumor size, CA-19-9 levels, and TNM stage. The ROC curve, with an AUC of 0.908 (p < 0.001), effectively distinguished CHOL tumor tissues from adjacent non-tumor tissues. And its sensitivity and specificity in distinguishing CHOL tissues from normal tissues were 88.5% and 89.2%, respectively. Survival analyses linked LUCAT1 overexpression to poorer patient outcomes. Silencing LUCAT1 impaired the proliferation and migration of QBC939 and HuCCT1 cells. Dual-luciferase assay confirmed the regulatory relationship between miR-141-3p and LUCAT1. Inhibition of miR-141-3p reversed the effect of LUCAT1 on the proliferation and migration of QBC939 and HuCCT1 cells. Conclusion LUCAT1 demonstrates significant diagnostic and prognostic potential and could serve as a novel biomarker for CHOL.

WDR62 affects the progression of ovarian cancer by regulating the cell cycle

Abstract Background Ovarian Cancer (OC) is a gynecological malignant tumor with an extremely high mortality rate, seriously endangering women’s health. Due to its insidious clinical manifestations, most patients are diagnosed in the advanced stage of the disease. The currently clinically relied CA125 has limited specificity for the early diagnosis of ovarian cancer. Hence, identifying new promising biomarkers is crucial for the early screening, diagnosis, and treatment of ovarian cancer. Based on differential expression analysis, WGCNA and survival analysis, we identified a centromere-associated gene, WDR62, which is highly expressed in ovarian cancer and highly correlated with ovarian cancer, as well as the poor prognosis of ovarian cancer patients with high expression, suggesting that WDR62 may be a potential biomarker for ovarian cancer. Previous studies have shown that WDR62 is closely associated with the occurrence, development and prognosis of a variety of tumors. However, its role in ovarian cancer has not been studied in depth. Methods Using combined TCGA and GTEx datasets from the UCSC database, along with WGCNA, and survival analysis, WDR62 was identified as a potential biomarker. GEPIA2 database, GEO database, qRT-PCR, and Western blot proved the expression of WDR62. Enrichment analysis, cell transfection, Western blots and CCK8 demonstrated the regulatory mechanism of WDR62, and the detailed mechanism of WDR62 involvement in the occurrence and development of ovarian cancer was predicted by interaction analysis and correlation analysis. Results WDR62 was highly expressed in ovarian cancer cells compared to normal ovarian epithelial cells, both at the RNA and protein levels. Patients with high WDR62 expression had a poor survival prognosis. Upon WDR62 knockdown, the expression of cell cycle-related proteins CDK1 and C-Myc decreased in ovarian cancer cells, and the cell proliferative capacity was decreased. Based on bioinformatic analysis, it was hypothesized that WDR62 might mediate the JNK signaling pathway by interacting with MAPK8, thus affecting ovarian cancer progression through cell cycle regulation. Conclusions WDR62 is overexpressed in ovarian cancer and is closely related to the prognosis of ovarian cancer patients. WDR62 promotes ovarian cancer progression by regulating the cell cycle and may influence its development through interaction with MAPK8 to mediate the JNK signaling pathway. These findings suggest that WDR62 could be a potential target for the early screening, diagnosis, and treatment of ovarian cancer.

Enhancer of mRNA Decapping protein 4 (EDC4) interacts with replication protein a (RPA) and contributes to Cisplatin resistance in cervical Cancer by alleviating DNA damage

Abstract Background Cervical cancer (CC) is the third most common gynecological malignancy around the world. Cisplatin is an effective drug, but cisplatin resistance is a vital factor limiting the clinical usage of cisplatin. Enhancer of mRNA decapping protein 4 (EDC4) is a known regulator of mRNA decapping, which was related with genome stability and sensitivity of drugs. This research was to investigate the mechanism of EDC4 on cisplatin resistance in CC. Two human cervical cancer cell lines, HeLa and SiHa, were used to investigate the role of EDC4 on cisplatin resistance in vitro. The knockdown or overexpression of EDC4 or replication protein A (RPA) in HeLa or SiHa cells was performed by transfection. Cell viability was analyzed by MTT assay. The growth of cancer cells was evaluated by colony formation assay. DNA damage was measured by γH2AX (a sensitive DNA damage response marker) immunofluorescent staining. The binding of EDC4 and RPA was analyzed by immunoprecipitation. Results EDC4 knockdown in cervical cancer cells (HeLa and SiHa) enhanced cisplatin sensitivity and cisplatin induced cell growth inhibition and DNA damage. EDC4 overexpression reduced DNA damage caused by cisplatin and enhanced cell growth of cervical cancer cells. EDC4 could interact with RPA and promote RPA phosphorylation. RPA knockdown reversed the inhibitory effect of EDC4 on cisplatin-induced DNA damage. Conclusion The present results indicated that EDC4 is responsible for the cisplatin resistance partly through interacting with RPA in cervical cancer by alleviating DNA damage. This study indicated that EDC4 or RPA may be novel targets to combat chemotherapy resistance in cervical cancer. Graphical abstract