Critical Reviews in Eukaryotic Gene Expression

RBM15-Mediated N6-Methyl Adenosine (m6A) Modification of EZH2 Drives the Epithelial-Mesenchymal Transition of Cervical Cancer

RBM15 functions as an oncogene in multi-type cancers. However, the reports on the roles of RBM15 in cervical cancer are limited. The purpose of this study was to investigate the potentials of RBM15 in cervical cancer. RT-qPCR was conducted to determine mRNA levels. Western was carried out to detect protein expression. CCK-8, colony formation and EdU assays were conducted to determine cell proliferation. Scratch and transwell assays were conducted to determine cell migration and invasion. MeRIP assay was conducted to determine N6-methyl adenosine (m6A) levels. Luciferase assay was conducted to verify the m6A sites of EZH2 and binding sites between EZH2 and promoter of FN1. ChIP assay was conducted to verify the interaction between EZH2 and FN1. The results showed that RBM15 was upregulated in cervical cancer patients and cells. Moreover, high levels of RBM15 predicted poor clinical outcomes. RBM15 knockdown inhibited the proliferation and epithelial-mesenchymal transition (EMT) of cervical cancer cells. RBM15 promoted the m6A modification of EZH2 as well as its protein translation. Additionally, EZH2 bound to the promoter of fibronectin 1 (FN1) and EZH2-FN1 axis is the cascade downstream of RBM15. Overexpressed EZH2 antagonized the effects of RBM15 knockdown and promoted the aggressiveness of cervical cancer cells. In summary, RBM15/EZH2/FN1 signaling cascade induces the proliferation and EMT of cervical cancer. Therefore, RBM15/EZH2/FN1 signaling may be a promising strategy for cervical cancer.

Foxm1-Mediated Transcriptional Inactivation of NLRP3 Inflammasome Promotes Immunosuppression in Cervical Cancer

Foxm1 functions as an oncogene in multiple human malignancies, including cervical cancer. However, the potential of Foxm1 in the tumor microenvironment (TME) is still unknown. The purpose of the present study is to investigate the role of Foxm1 in CD8+ T cell anti-tumor immunity. RT-qPCR is conducted to calculate mRNA levels. JASPAR is used to predict the binding sites between Foxm1 and NLRP3. ChIP assay is performed to verify the occupancy of Foxm1 on the promoter of NLRP3. Modulatory relationship between Foxm1 and NLRP3 is verified by luciferase assay. <i>In vivo</i> assays are conducted to further verify the role of Foxm1/NLRP3 axis in cervical cancer. HE staining assay is applied for histological analysis. Flow cytometry is conducted to determine the functions of immune cells. We found that Foxm1 knockdown decreases tumor burden and suppresses tumor growth of cervical cancer. Foxm1 knock-down promotes the infiltration of CD8+ T cells. Foxm1 deficiency inhibits the exhaustion of CD8+ T cells and facilitates the maintenance of CD8+ effector and stem-like T cells. Moreover, Foxm1 transcriptionally inactivates NLRP3 and suppresses the expression of innate cytokines IL-1β and IL-18. However, inhibition of NLRP3 inflammasome or neutralizing IL-1β and IL-18 inhibits anti-tumor immunity and promoted tumor growth in Foxm1 deficiency in CD8+ T cells. In summary, targeting Foxm1 mediates the activation of NLRP3 inflammasome and stimulates CD8+ T cell anti-tumor immunity in cervical cancer.

HDAC1-Mediated Downregulation of NEU1 Exacerbates the Aggressiveness of Cervical Cancer

HDAC1 functions as an oncogene in multi-type cancers. This study aimed to investigate the roles of histone deacetylase 1 (HDAC1) in cervical cancer (CC). mRNA expression was determined using reverse transcription quantitative polymerase chain reaction. The protein-protein complexes was analyzed using co-immunoprecipitation assay. The binding sites between NRF2 and NEU1 were confirmed by chromatin immunoprecipitation assay. Cell viability was detected by CCK-8. Cell proliferation was measured using CCK-8 and colony formation assays. Cell migrative and invasive ability were determined using transwell assay. We found that HDAC1 was upregulated in CC patients and cells. Trichostatin A (TSA) treatment decreased the number of colonies and migrated and invaded cells. Moreover, HDAC1 interacted with NRF2 to downregulate NEU1 expression. NEU1 knockdown attenuated the effects of TSA and enhanced the aggressiveness of CC cells. In conclusion, HDAC1 functions as an oncogene in CC. Targeting HDAC1 may be an alternative strategy for CC.

Analysis of the Secreted Peptidome from Omental Adipose Tissue in High-Grade Serous Ovarian Cancer

High-grade serous ovarian cancer (HGSOC) is a preferential omental metastasis malignancy. Since omental adipose tissue is an endocrine organ, we used liquid chromatography tandem mass spectrometry (LC-MS/MS) to compare the peptides secreted from omental adipose tissues of HGSOC and benign serous ovarian cysts (BSOC). Among the differentially secreted peptides, we detected 58 upregulated peptides, 197 downregulated peptides, 24 peptides that were only in the HGSOC group and 20 peptides that were only in the BSOC group (absolute fold change ≥ 2 and <i>P</i> < 0.05). Then, the basic characteristics of the differential peptides were analyzed, such as lengths, molecular weights, isoelectric points, and cleavage sites. Furthermore, we summarized the possible functions according to the precursor protein functions of the differentially expressed peptides by Gene Ontology (GO) analysis with the Annotation, Visualization, and Integrated Discovery (DAVID) database and canonical pathway analysis with IPA. For the GO analysis, the differentially secreted peptides were mainly associated with binding in molecular function and cellular processes in biology process. For the canonical pathways, the differentially secreted peptides were related to calcium signaling, protein kinase A signaling, and integrin-linked kinase (ILK) signaling. We also identified 67 differentially secreted peptides that located in the functional domains of the precursor proteins. These functional domains were mainly related to energy metabolism and immunoregulation. Our study might provide drugs that could potentially treat HGSOC or omental metastases of HGSOC cells.

lncRNA799/TBL1XR1/ZEB1 Axis Forms a Feedback Loop to Promote the Epithelial-Mesenchymal Transition of Cervical Cancer Cells

Cervical cancer is a common malignancy among women worldwide. Long non-coding RNAs (lncRNAs) are frequently involved in the pathogenesis of cervical cancer. Therefore, the present study aimed to investigate the potentials of lncRNA799 in cervical cancer. mRNA and protein expression were detected by reverse transcription-quantitative polymerase chain reaction and Western blot analysis, respectively. Cellular functions were assessed using CCK-8, wound healing and transwell analysis. The binding potential of zinc finger E-box-binding homeobox 1 (ZEB1) on the promoter of lncRNA799 was predicted utilizing the JASPAR database, and was then verified by luciferase and chromatin immunoprecipitation (ChIP) assays. Furthermore, the gene interactions were assessed using RNA immunoprecipitation and co-immunoprecipitation assays. The results demonstrated that lncRNA799 was upregulated in cervical cancer cells. However, lncRNA799 deficiency suppressed the proliferation and epithelial-mesenchymal transition of cervical cancer cells. Furthermore, lncRNA799 could interact with eukaryotic translation initiation factor 4A3 to maintain the mRNA stability of transducin (β)-like 1 X-linked receptor 1 (TBL1XR1) and promote the interaction between ZEB1 and TBL1XR1. Additionally, the results showed that ZEB1 could transcriptionally activate lncRNA799. Taken together, the present study suggested that the lncRNA799/TBL1XR1/ZEB1 axis could form a positive feedback loop in cervical cancer and could be, therefore, considered as a potential therapeutic strategy for cervical cancer.

A Five-Gene Expression Signature Predicts Ovarian Cancer Metastasis

Ovarian cancer represents one of the most malignant gynecological tumors. Despite recent advances in treatment, ovarian cancer remains to be highly susceptible to metastasis. However, information concerning genome-wide gene expression profiles is limited to develop a metastasis-specific gene signature in ovarian cancer. In this work, we try to identify changes in gene expression profile that underlie ovarian cancer metastasis. The dataset GSE73168 deposited in the Gene Expression Omnibus (GEO) database was processed to identify differentially expressed genes (DEGs) between primary tumor and metastatic tumor samples. The weighted gene correlation network analysis (WGCNA) was conducted for modules related to ovarian cancer metastasis. Modular genes associated with ovarian cancer metastasis were summarized for the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Receiver operating characteristic (ROC) curves were plotted to estimate the superiority of candidate genes in detecting ovarian cancer metastasis. The WGCNA yielded 25 co-expression network modules in the dataset GSE73168, and highly correlated genes with ovarian cancer metastasis were identified in the blue module. Twenty-two genes demonstrated differential expression between primary tumor and metastatic tumor samples, and two downregulated genes (P2RY13 and NKX6-1) and three upregulated genes (CD36, LOC57399 and RP11-587D21.4) of these 22 DEGs was also shown to correlate with ovarian cancer metastasis in the blue module. The area under the ROC curve verified these five DEGs as metastasis-specific genes for ovarian cancer. These results show P2RY13, NKX6-1, CD36, LOC57399 and RP11-587D21.4 serve as metastasis-specific genes for ovarian cancer.

MMP-16 as a New Biomarker for Predicting Prognosis and Chemosensitivity of Serous Ovarian Cancer: A Study Based on Bioinformatics Analysis

To explore the prognostic value of MMP-16 expression in patients with serous ovarian cancer and the usefulness of MMP-16 expression to predict sensitivity to chemoradiotherapy. The relationship between MMP-16 expression and clinicopathological parameters of serous ovarian cancer was evaluated in The Cancer Genome Atlas (TCGA) database. Cox proportional hazard regression analysis was performed to measure the prognostic significance of MMP-16 in serous ovarian cancer. Dataset GSE51373 was applied to estimate the difference of MMP-16 expression between chemotherapy-sensitive group and resistant group of serous ovarian cancer. Receiver operating characteristic (ROC) curve was also drawn. In addition, the online tool Kaplan-Meier Plotter was used to assess the prognostic value of MMP-16 in patients with serous ovarian cancer. A total of 235 patients with serous ovarian cancer were included in the TCGA database. Cox regression univariate analysis showed that high expression of MMP-16 was not conducive to the overall survival of patients with serous ovarian cancer (hazard ratio [HR] = 1.47, 95% CI: 1.03~2.08; P < 0.05). The results of Cox regression multivariate analysis also demonstrated that there was a statistically significant difference. The results of the online database Kaplan-Meier Plotter analysis showed that the high expression of MMP-16 was not conducive to the progression-free survival (PFS) of patients with serous ovarian cancer (HR = 1.26, 95% CI: 1.06~1.29; P < 0.05). The expression of MMP-16 in the chemotherapy-sensitive group was notably lower than that in the chemotherapy-resistant group, which had a moderate predictive value in predicting the chemosensitivity of serous ovarian cancer (AUC = 0.7187). High expression of MMP-16 is not conducive to chemotherapy sensitivity and survival of patients with serous ovarian cancer, and has predictive value for chemotherapy resistance and prognosis.

The Role of Lysophosphatidic Acid Receptors in Ovarian Cancer: A Minireview

Lysophosphatidic acid (LPA) is a bioactive lipid component of ovarian cancer activating factor, which is present at a high concentration in the ascitic fluid and plasma of patients with ovarian cancer. A group of six lysophosphatidic acid receptors (LPARs), LPAR1 through LPAR6, which belong to the G protein-coupled receptor superfamily (GPCR), mediate cellular activities of LPA and activates a series of downstream molecules and cellular responses, including biological and pathological effects. LPARs are widely expressed in normal ovary, benign tumor, and ovarian cancer tissues and cancer cell lines with a broad range of levels. The LPA/LPAR axis is involved in tumorigenesis and development of ovarian cancer through mediating the cellular responses to LPA and influencing the expression and function of oncogenic molecules. In the present review, the roles of LPARs in ovarian cancer, including the expression, function, and downstream molecules, are summarized, and we discuss the implications for ovarian cancer treatment that targets LPARs.

IncRNA TRAF3IP2-AS1 Restrains Cervical Cancer Progression by Sponging miR-3677-3p and Acts As a Diagnostic Marker in Cervical Cancer

Long non-coding RNAs (lncRNAs) are important participants in human tumors. The current study assessed the clinical value and functional role of TRAF3IP2-AS1 in cervical cancer. Quantitative reverse transcription polymerase chain reaction was used to compare the TRAF3IP2-AS1 levels in serum samples in healthy control, patients with cervical cancer and cervical intraepithelial neoplasia (CIN), as well as in cervical cancer cells. Receiver operator characteristic curve analysis explored the potential diagnostic potential of TRAF3IP2-AS1. Functional and rescue experiments were carried out for analysis of cellular behaviors and miRNA interplays. TRAF3IP2-AS1 was discovered to be downregulated in the serum of cervical cancer patients and could distinguish cervical cancer patients from CIN and healthy individuals. The elevated TRAF3IP2-AS1 expression restrained cell proliferation, migration, and invasion. TRAF3IP2-AS1 could bind with miR-3677-3p to regulate cervical cancer cellular behaviors. TRAF3IP2-AS1 may play a tumor-suppressor role in cervical cancer progression by sponging miR-3677-3p. TRAF3IP2-AS1 appears to have a potential diagnostic value and be a promising treatment target for treating cervical cancer.

CCT3 as a Diagnostic and Prognostic Biomarker in Cervical Cancer

The chaperonin-containing TCP1 complex subunit 3 (CCT3) has been reported to be involved in the development and prognosis of many tumors, including cervical cancer (CC). This study aimed to analyze the expression and prognostic value of CCT3 in CC by bioinformatics and retrospective study. CCT3 gene expression profiles and clinical information in CC were downloaded from the cancer genome atlas (TCGA) and gene expression omnibus (GEO) databases. CCT3 expression was verified by quantitative real-time polymerase chain reaction (RT-qPCR), Western blot, and immunohistochemistry (IHC). Logistic regression and chi-square testing were used to analyze the relationship between CCT3 expression and the clinical characteristics of CC. Kaplan-Meier and Cox analyses were used to evaluate whether CCT3 affects the prognosis of CC. Nomogram and calibration curves were used to test the predictive value of CCT3. The expression of CCT3 in CC tissues was significantly upregulated compared with that in adjacent benign tissues, and was related to HPV16/18 infection, grade, and positive lymph nodes. High expression of CCT3 is associated with poor prognosis of CC and can be used as an independent risk factor for CC. The prognostic model based on CCT3 and CC clinical features has good predictive ability. CCT3 is overexpressed in CC, which is related to poor prognosis and expected to become a biomarker for CC.

Palmdelphin Inhibits Ovarian Cancer Cell Stem Specification via Downregulating Ring Finger Protein 145

This study aimed to investigate the roles of PALMD in ovarian cancer. mRNA expression was detected using RT-qPCR. The aldehyde dehydrogenase (ALDH) activity and biomarkers of ovarian cancer stem cells were determined using flow cytometry. The stemness of ovarian cancer cells was determined using sphere formation assay. Cell viability was determined using CCK-8 assay. The number of colonies was determined using colony formation assay. Cell migration was detected using wound healing assay. Cell invasion was determined using transwell assay. The results showed that PALMD was downregulated in ovarian cancer. Overexpressed PALMD inhibited the proliferative, migrative, and invasive ability of ovarian cancer cells. Moreover, PALMD inhibited the stem-like properties of ovarian cancer cells. Additionally, PALMD downregulated ring finger protein 145 (RNF145) expression, overexpression of which contributed to the aggressiveness of ovarian cancer cells. Taken together, PALMD suppressed ovarian cancer cell stem specification via inhibiting RNF145 expression.

TRIP13 Is a Potential Prognostic Marker and Therapeutic Target for Endometrial Cancer

Uterine corpus endometrial carcinoma (UCEC) is a prevalent malignancy within the female reproductive system, with a rising global incidence. Although thyroid hormone receptor interacting protein 13 (TRIP13) has been implicated in various tumor etiologies and progressions, its role in UCEC remains poorly characterized. This study aimed to delineate TRIP13's expression profile in UCEC by analyzing transcriptome data from multiple databases. We investigated genomic alterations and epigenetic modifications of the TRIP13 gene using the cBioPortal tool. The prognostic value of TRIP13 was assessed via Kaplan-Meier survival analysis and Cox regression modeling. Additionally, we examined TRIP13's impact on immunotherapy responsiveness and chemotherapy sensitivity through immunological and pharmacological analyses. The expression of TRIP13 in both normal endometrial and cancer cell lines was evaluated using quantitative real-time polymerase chain reaction (qPCR). Our findings reveal that TRIP13 expression in UCEC tumor samples is significantly higher than in normal tissues and increases with tumor grade and stage progression. High TRIP13 expression is significantly associated with poor prognosis in UCEC patients, establishing it as an independent prognostic biomarker. TRIP13 shows a positive correlation with immunosuppressive cell infiltration and a negative correlation with immune-activating cell infiltration, suggesting a potential role in tumor immune evasion. Further analysis identified TRIP13 as a potential biomarker for predicting immunotherapy response. Moreover, TRIP13 expression is significantly associated with sensitivity to certain chemotherapeutic agents, indicating its potential as a therapeutic target. qPCR experiments confirmed the overexpression of TRIP13 in endometrial cancer cell lines. The role of TRIP13 in modulating the tumor immune microenvironment, as well as its predictive value for immunotherapy and chemotherapy responses, underscores its importance in developing personalized treatment strategies for UCEC. These findings provide novel molecular targets and therapeutic insights for a precision medicine approach to UCEC.

Epigenetic Signatures and Prognostic Biomarkers Analysis of Methylation-Driven Genes in Uterine Endometrial Carcinosarcoma

Uterine corpus endometrial carcinoma (UCEC) is one of the most common gynecological malignancies, and understanding the molecular mechanisms underlying its development is essential for improving diagnosis and treatment. However, the role of DNA methylation, a key epigenetic modification, in UCEC prognosis prediction and clinical treatment strategies has rarely been studied. This study utilized publicly available datasets from The Cancer Genome Atlas (TCGA) and online bioinformatics tools to analyze the differential methylation and expression of six selected genes: TP53, PTEN, PTX3, TNK1, PPP2R1A, and KLRG2. These genes were chosen based on their known roles in cancer-related pathways, previous associations with oncogenic processes, and preliminary data showing significant changes in methylation and expression in UCEC compared with normal tissues. We integrated mRNA expression and DNA methylation data with the MethylMix method to identify genes with methylation-driven expression changes. Our analysis revealed that these genes exhibit distinct differential expression and methylation patterns in UCEC, suggesting potential regulatory mechanisms. The expression patterns across the six genes were observed, and TP53, TNK1, PPP2R1A, and KLRG2 were upregulated in tumors, and PTX3 was downregulated in tumors. At the same time, there was no significant change in the expression of PTEN gene. The differential expression correlates with changes in methylation, providing insights into the gene regulation occurring in UCEC. Additionally, Kaplan-Meier survival analysis revealed that the expression levels of specific genes, particularly PTX3, TNK1, and KLRG1, are significantly associated with overall survival in UCEC patients. Higher expression of these genes correlated with poorer survival outcomes, suggesting their potential as prognostic markers. In contrast, the expression of TP53, PTEN, and PPP2R1A did not show a significant impact on patient survival. The functional importance of these genes was investigated utilizing pathway enrichment and protein-protein interaction networks. Additionally, pathway enrichment analysis indicated these genes are involved in critical cancer pathways. The findings highlight the importance of integrating epigenetic and transcriptomic data to understand UCEC pathogenesis and suggest that the identified genes could serve as potential biomarkers for early diagnosis and treatment strategies.

circRNA circRIMS Downregulates miR-505 through Methylation to Suppress Cell Proliferation in Endometrial Cancer

It is known that the circular RNA (circRNA) molecule circRIMS is overexpressed in gastric cancer and plays an oncogenic role. However, its role in other cancers is unknown. In this study, we analyzed its role in endometrial cancer (EC). EC and paired non-tumor tissue samples were collected from a total of 63 EC patients and subjected to total RNA isolations and reverse transcription quantitative polymerase chain reaction (RT-qPCR) to analyze the differential expression of circRIMS and miR-505. Overexpression of circRIMS and miR-505 was reached in EC cells and their interaction was analyzed using RT-qPCRs. The role of circRIMS in regulating miR-505 methylation was analyzed by methylation-specific RT-qPCR. Bromodeoxyuridine (BrdU) assay was performed to analyze the roles of circRIMS and miR-505 in regulating cell proliferation. circRIMS was upregulated in EC, while miR-505 was downregulated in EC. circRIMS and miR-505 were inversely correlated across both EC and non-tumor tissues. In EC cells, circRIMS overexpression decreased miR-505 expression and increased miR-505 gene methylation. BrdU assay showed that circRIMS overexpression increased cell proliferation and reduced the inhibitory effects of miR-505 overexpression on cell proliferation. circRIMS may downregulate miR-505 through methylation to increase cell proliferation.

LncRNA FOXCUT Stimulates the Progression of Endometrial Cancer

Endometrial cancer (EC) is a common gynecological tumor and the third leading cause of cancer-related death in women. Previous research has proved that long noncoding RNA (lncRNA) FOXCUT acts as an oncogene in several cancers. This study ascertained that lncRNA FOXCUT was highly expressed in endometrial cancer cells. Overexpression of lncRNA FOXCUT promoted EC cell proliferation, invasion, and migration, as well as epithelial-mesenchymal transition (EMT). In addition, overexpression of lncRNA FOXCUT could inhibit the apoptosis of cancer cells and block EC cells in S phase, whereas silencing lncRNA FOXCUT had the opposite effect. Thus, lncRNA FOXCUT had a regulatory effect on promoting the progression of EC cells, which might provide novel potential targets for research on targeted therapy of EC.

MiR-18a-5p Promotes Proliferation, Migration, and Invasion of Endometrial Cancer Cells by Targeting THBD

The purpose of this study was to elucidate the role that the miR-18a-5p/THBD regulatory pathway plays in endometrial cancer (EC), which could provide a theoretical basis for potential therapeutic targets. Differentially expressed genes in EC tissue and normal tissue were determined by bioinformatics analysis. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to compare the expression of miR-18a-5p and THBD mRNA in normal human endometrial cells and human EC cells. CCK-8 assay was used to compare the proliferative ability of EC cells in different treatment groups. Transwell assay was used to detect the migratory and invasive abilities of EC cells in different treatment groups. Dual-luciferase assay was used to verify the targeting relationship between miR-18a-5p and THBD. Western blot assay was used to detect THBD protein expression level. qRT-PCR results showed that miR-18a-5p was significantly upregulated in EC cells, and expression of its target gene, THBD, was significantly downregulated. CCK-8 and transwell assays showed that miR-18a-5p could enhance the proliferative, migratory, and invasive abilities of EC cells, whereas THBD could weaken those abilities. Dual-luciferase assay confirmed that miR-18a-5p could negatively regulate THBD expression. In addition, rescue experiments revealed that the oncogenic effect of miR-18a-5p on EC cells was inhibited by THBD overexpression. We conclude that miR-18a-5p could promote the proliferation, migration, and invasion of EC cells by targeting and downregulating THBD expression, and the miR-18a-5p/THBD regulatory pathway might be a therapeutic target. The results of this study may serve as a theoretical basis for related drug development.