Archives of Biochemistry and Biophysics

Fatostatin inhibits the development of endometrial carcinoma in endometrial carcinoma cells and a xenograft model by targeting lipid metabolism

Endometrial carcinoma is a type of gynecological cancer that originates in the endometrial epithelial tissue. Due to its high proliferation and ability to invade muscle tissue, it is one of the most common malignant tumors in the female reproductive system. Fatostatin is a small molecule non-sterol diarylthiazole derivative that acts as a chemical inhibitor of the sterol regulatory-element binding protein (SREBP) pathway. Previous studies have shown that fatostatin has an anti-tumor effect in some cancers. In this study, we investigated the effect of fatostatin on the growth, proliferation, apoptosis, migration and cell cycle of human endometrial carcinoma cells (HEC-1A and AN3 CA cells) using cholecystokinin (CCK) -8 method, clonogenicity assay, wound closure assay, Transwell migration assay and flow cytometer. We also examined its effect on the expression of apoptosis-associated protein (Caspase-3, Caspase-8 and Caspase-9) and level of lipid metabolism-related proteins, free fatty acid, and total cholesterol in cells. The growth of endometrial carcinoma xenografts was measured to confirm the effect of fatostatin in vivo. Our results showed that fatostatin inhibited the growth and proliferation of human endometrial carcinoma cells, changed their cell cycle and induced apoptosis. Based on the preliminary animal experiments, fatostatin also exhibited antitumor activity. The present study adds a new dimension to our understanding of the antitumor effects of fatostatin and provides an experimental basis for its use, and supports its potential value for clinical application.

RNF113A as a poor prognostic factor promotes metastasis and invasion of cervical cancer through miR197/PRP19/P38MAPK signaling pathway

It has been discovered that aberrant expression of RNF113A plays a significant role in various diseases, including esophageal cancer, hepatocellular carcinoma, and X-linked trichothiodystrophy syndrome. Nevertheless, its functional implications in cervical cancer (CC) remain unclear. The objective of this study was to investigate the role of RNF113A in both the development and prognosis of CC. To achieve this objective, a total of sixty cases were included in the follow-up investigation. The findings revealed a significant up-regulation of RNF113A protein in CC tissues compared to paired paracancerous tissues, and a high expression level of RNF113A was strongly associated with malignant phenotypes such as lymph node metastasis, differentiation degree, depth of invasion, and FIGO stage. Meanwhile, RNF113A was found to be an independent prognostic risk factor, with its high expression significantly correlating with a reduced overall survival period in patients. To elucidate the underlying cause and mechanism of the unfavorable prognosis associated with RNF113A, comprehensive functional investigations were conducted both in vitro and in vivo.Interestingly, it was revealed that RNF113A promoted migration and invasion while inhibiting apoptosis of CC cells, thereby contributing to a poor prognosis. Mechanistically, RNF113A regulated the progression and prognosis of CC through the miR197/Prp19/p38Mark signaling pathway. Overall, our findings underscore the potential clinical significance of RNF113A as an unfavorable prognostic factor in CC.

Promotive actions of lncRNA EBLN3P involved in cervical cancer progression via interacting with miR-29c-3p and TAF15 to modify RCC2

Cervical cancer is a common cancer that seriously affects women's health globally. The key roles of long non-coding RNAs (lncRNAs) in the onset and development of cervical cancer have attracted much attention. Our study aims to uncover the roles of lncRNA EBLN3P and miR-29c-3p and the mechanisms by which EBLN3P and miR-29c-3p regulate malignancy in cervical cancer. Tumor and adjacent normal tissues were collected from cervical cancer patients, and the expression of EBLN3P and miR-29c-3p were analyzed via RT-qPCR. The capacities of proliferation, migration, and invasion were assessed using CCK-8, wound healing and transwell assays. The interaction among EBLN3P, miR-29c-3p and TAF15 was determined by luciferase, RNA immunoprecipitation and RNA pull-down assays, respectively. A subcutaneous tumor xenograft mouse model was established to evaluate the functional role of EBLN3P in vivo. The interaction and reciprocal negative regulation between EBLN3P and miR-29c-3p were uncovered in cervical cancer cells. Likewise, EBLN3P and miR-29c-3p expression patterns in tumor tissues presented a negative association. EBLN3P knockdown weakened cell proliferation, migration and invasion, but these effects were abrogated by miR-29c-3p depletion. Mechanistically, ALKBH5 might impaired EBLN3P stability to reduce its expression. EBLN3P functioned as a competing endogenous RNA (ceRNA) for miR-29c-3p to relieve its suppression of RCC2. Besides, EBLN3P enhanced RCC2 mRNA stability via interacting with TAF15. Furthermore, silencing of EBLN3P repressed the tumor growth in mice. Altogether, lncRNA EBLN3P positively regulates RCC2 expression via competitively binding to miR-29c-3p and interacting with TAF15, thereby boosting proliferation, migration, and invasion of cervical cancer cells.

Advances in molecular mechanism of HPV16 E5 oncoprotein carcinogenesis

For a considerable duration, cervical cancer has posed a significant risk to the well-being and survival of women. The emergence and progression of cervical cancer have garnered extensive attention, with prolonged chronic infection of HPV serving as a crucial etiological factor. Consequently, investigating the molecular mechanism underlying HPV-induced cervical cancer has become a prominent research area. The HPV molecule is composed of a long control region (LCR), an early coding region and a late coding region.The early coding region encompasses E1, E2, E4, E5, E6, E7, while the late coding region comprises L1 and L2 ORF.The investigation into the molecular structure and function of HPV has garnered significant attention, with the aim of elucidating the carcinogenic mechanism of HPV and identifying potential targets for the treatment of cervical cancer. Research has demonstrated that the HPV gene and its encoded protein play a crucial role in the invasion and malignant transformation of host cells. Consequently, understanding the function of HPV oncoprotein is of paramount importance in comprehending the pathogenesis of cervical cancer. E6 and E7, the primary HPV oncogenic proteins, have been the subject of extensive study. Moreover, a number of contemporary investigations have demonstrated the significant involvement of HPV16 E5 oncoprotein in the malignant conversion of healthy cells through its regulation of cell proliferation, differentiation, and apoptosis via diverse pathways, albeit the precise molecular mechanism remains unclear. This manuscript aims to provide a comprehensive account of the molecular structure and life cycle of HPV.The HPV E5 oncoprotein mechanism modulates cellular processes such as proliferation, differentiation, apoptosis, and energy metabolism through its interaction with cell growth factor receptors and other cellular proteins. This mechanism is crucial for the survival, adhesion, migration, and invasion of tumor cells in the early stages of carcinogenesis. Recent studies have identified the HPV E5 oncoprotein as a promising therapeutic target for early-stage cervical cancer, thus offering a novel approach for treatment.

Tanshinone IIA inhibits endometrial carcinoma growth through the MAPK/ERK/TRIB3 pathway

Endometrial carcinoma is the most common gynecological tumor in developed countries. Tanshinone IIA is a traditional herbal medicine which is to treat cardiovascular disease and has been shown to have various biological effects, such as anti-inflammatory, antioxidative and antitumor activities. However, there has been no study about the effect of tanshinone IIA on endometrial carcinoma. Thus, the aim of this study was to determine the antitumor activity of tanshinone IIA against endometrial carcinoma and investigate the associated molecular mechanism. We demonstrated that tanshinone IIA induced cell apoptosis and inhibited migration. We further demonstrated that tanshinone IIA activated the intrinsic (mitochondrial) apoptotic pathway. Mechanistically, tanshinone IIA induced apoptosis by upregulating TRIB3 expression and inhibiting the MAPK/ERK signaling pathway. In addition, knockdown of TRIB3 with an shRNA lentivirus accelerated proliferation and attenuated inhibition mediated by tanshinone IIA. Finally, we further demonstrated that tanshinone IIA inhibited tumor growth by inducing TRIB3 expression in vivo. In conclusion, these findings suggest that tanshinone IIA has a significant antitumor effect by inducing apoptosis and may be used as a drug for the treatment of endometrial carcinoma.

20(S)-Rg3 upregulates FDFT1 via reducing miR-4425 to inhibit ovarian cancer progression

We previously found that ginsenoside 20(S)-Rg3 diminishes the proliferative and invasive capacities of ovarian cancer cells by decreasing miR-4425 level. Yet the mechanism of action of miR-4425 in ovarian cancer remains unclear. Here we report that miR-4425 is upregulated in ovarian cancer tissues relative to normal ovarian tissues, and transfection of miR-4425 inhibitor impairs the proliferation, migration and invasion of SKOV3 and 3AO ovarian cancer cells. Further, miR-4425 antagomiR reduces cell proliferation in a subcutaneous SKOV3 xenograft model using BALB/c nude mice. We identifies farnesyl-diphosphate farnesyltransferase 1 (FDFT1) as a direct target of miR-4425 by Western blotting and a luciferase reporter assay. Forced expression of FDFT1 via transfection of an FDFT1-expressing plasmid into ovarian cancer cells not only retards cell proliferation, motility and invasiveness, but also negates the tumorigenic properties of a miR-4425 mimic. By contrast, silencing of FDFT1 by siRNAs abrogates suppression of the proliferation, migration and invasion of ovarian cancer cells treated with a miR-4425 inhibitor. Finally, transfection of either a miR-4425 mimic or FDFT1 siRNAs into 20(S)-Rg3-treated ovarian cancer cells counteracts the tumor-inhibitory activity of the ginsenoside. In conclusion, 20(S)-Rg3 exerts anti-ovarian cancer activity by downregulating oncogenic miR-4425 that inhibits the expression of the tumor suppressor gene FDFT1. These results expand our current understanding of the molecular pathways leading to ovarian cancer progression, and unveil the mechanism of action of 20(S)-Rg3 in ovarian cancer inhibition.

IL-6 and IL-8 are involved in JMJD2A-regulated malignancy of ovarian cancer cells

Emerging evidence shows that histone modification and its related regulators are involved in the progression and chemoresistance of ovarian cancer (OC) cells. Our present study found that the expression of Jumonji C domain-containing 2A (JMJD2A), while not JMJD2B or JMJD2C, is increased in OC cells and tissues as compared with that in their corresponding controls. Knockdown of JMJD2A can decrease proliferation while increase cisplatin (CDDP) sensitivity of OC cells. By screening the expression of cytokines involved in the progression of ovarian cancer, we found that knockdown of JMJD2A can inhibit the expression of interleukin-6 (IL-6) and IL-8 in ovarian cancer cells. Recombinant IL-6 (rIL-6) and rIL-8 can attenuate si-JMJD2A-suppressed malignancy of OC cells. Mechanistically, JMJD2A can directly bind with the promoter of IL-6 to trigger its transcription. For IL-8, JMJD2A can increase it mRNA stability in OC cells. Collectively, we revealed that JMJD2A can trigger the malignancy of OC cells via upregulation of IL-6 and IL-8. It suggested that JMJD2A might be a potential target for OC treatment and therapy.

Quercetin attenuates metastatic ability of human metastatic ovarian cancer cells via modulating multiple signaling molecules involved in cell survival, proliferation, migration and adhesion

Ovarian cancer is the most deadly gynaecology related cancer due to its high metastasizing ability. Quercetin is the most abundant flavonoids received increased interest due to its anti-cancer properties. Although the anticancer property of quercetin is very well known, its anti-metastatic effect on metastatic ovarian cancer cells and their underlying molecular mechanism remains to be elucidated. Quercetin treatment at 50 μM and 75 μM concentration inhibit human metastatic ovarian cancer PA-1 cell survival and proliferation via inactivating PI3k/Akt, Ras/Raf pathways and EGFR expression. It also alters the expression of N-cadherin in PA-1 cells. Quercetin also decreases the secretion of gelatinase enzyme, proteolytic activity of MMP-2/-9, and both MMPs gene expression in metastatic ovarian cancer PA-1 cells. In addition to this quercetin inhibits the migration of PA-1 cells. Treatment of quercetin with PA-1 cells also downregulates the tight junctional molecules such as Claudin-4 and Claudin-11 while upregulates the expression of occludin. It is further validated by cell adhesion assay in which quercetin reduces the adhesion of PA-1 ovarian cancer cells. Results suggest that quercetin inhibits cell survival, proliferation, migration, and adhesion which plays crucial role in ovarian cancer metastasis. Hence, it could be a valuable therapeutic drug for the treatment and prevention of metastatic ovarian cancer.

Human papillomavirus 16 E6/E7 contributes to immune escape and progression of cervical cancer by regulating miR-142–5p/PD-L1 axis

Persistent infection of human papillomavirus (HPV) and immune escape are the main causes of cervical cancer. E6/E7 encoded by HPV16 may be closely related to carcinogenesis. HPV infection may upregulate PD-L1 expression, resulting in immune escape and cervical cancerigenesis. Evidence indicates that miRNAs may mediate the regulation of E6/E7 on PD-L1. Therefore, we aimed to screen the miRNA, and further verify its expression and functions. Bioinformatics approaches were used to screen the miRNAs that mediate the regulation of E6/E7 on PD-L1. The expression of the miRNA and PD-L1 in HPV

E6E7 regulates the HK2 expression in cervical cancer via GSK3β/FTO signal

Cervical cancer is one of the most common cancers in women worldwide. Hexokinase 2 (HK2) is responsible for phosphorylating glucose into glucose-6-phosphate, which is required for tumorigenesis and metastasis. E6E7 and FTO were exogenously expressed, and their effects on HK2 mRNA and protein levels were detected by RT-qPCR and Western blot. The exogenous expression of E6E7 in SiHa and C33A cells up-regulated the mRNA and protein levels of intracellular HK2, up-regulated the total m This study found that E6E7 oncogene activates the transcription of GSK3β; GSK3β can promote the ubiquitination-proteasomal degradation of FTO and reduce the level of FTO protein; FTO inhibits the maturation and translation of HK2 mRNA by retaining HK2 pre-mRNA in the nucleus.

Pharmacological USP2 targeting suppresses ovarian cancer growth by potentiating apoptosis and ferroptosis

Ovarian cancer is a frequently observed type of gynaecologic malignancy generally associated with poor prognosis around the world. Ubiquitin-specific proteases (USPs) form the largest subfamily of deubiquitylating enzymes and have emerged as potential therapeutic targets against human cancers. Through a systematic analysis of the prognostic significance of USP expression, USP2 was found to be inversely correlated with patient survival in ovarian cancer. Accordingly, we investigated the effects of pharmacological inhibition of USP2 on ovarian cancer by exploiting its small molecule inhibitor ML364. Our findings show that ML364 effectively hindered ovarian cancer growth and migration using a series of in vitro assays. In addition to apoptosis induction, ML364 also sensitized ovarian cancer cells to ferroptosis. Mechanistically, ML364 treatment resulted in cyclin D1 downregulation, increased poly (ADP-ribose) polymerase (PARP) cleavage, and elevated ROS levels in ovarian cancer cells. Collectively, our findings suggest USP2 as a potential therapeutic target in ovarian cancer, and hence, its pharmacological inhibition warrants further investigation.

Construction of a circRNA-miRNA-mRNA ceRNA regulatory network identifies RNAs and genes linked to human ovarian clear cell carcinoma

Ovarian clear cell carcinoma (OCCC) is an aggressive epithelial ovarian cancer subtype that prevalent in East Asia. Competing endogenous RNA (ceRNA) networks involving circular RNAs (circRNAs) and microRNAs (miRNAs) represent underexplored regulatory layers in OCCC pathogenesis. Multi-omics integration of circRNA (GSE271851 and GSE266248) and miRNA (GSE230956 and GSE200852) datasets from GEO was performed using DESeq2 and limma tools (|log2FC|≥1, p < 0.05). Experimentally validated circRNA-miRNA interactions were predicted using circBank 2.0, Circular RNA Interactome, and Starbase v3.0. ceRNA networks were constructed using inverse correlation principles and visualized in Cytoscape. Immune associations were assessed using TIMER2.0. Prognostic and expression validations were performed using the Kaplan-Meier Plotter and HPA databases, respectively. Three core circRNAs (hsa_circ_0002822, hsa_circ_0003641 and hsa_circ_0030509) were identified as being dysregulated across OCCC models. Their miRNA sponging activity formed a 65-node ceRNA network involving 15 miRNAs (e.g., miR-143-3p and miR-182-5p) and 47 mRNAs. Six targets (BRCA1, KRAS, MSH6, SMARCA4, SRC, and TSC1) exhibited significant correlations with immune infiltration (CD8 This study delineates a circRNA-driven ceRNA network in OCCC, and identifies BRCA1, KRAS, and MSH6 as multi-omic biomarkers with immune-modulatory roles. These findings provide a basis for the development of RNA-targeted therapies against this ovarian cancer subtype.

TRIM37 promotes the aggressiveness of ovarian cancer cells and increases c-Myc expression by binding to HUWE1

Tripartite motif containing 37 (TRIM37) was reported to function as a tumor promoter in the development of various cancers. However, the biological role of TRIM37 in ovarian cancer (OC) was still unclear. Expressions of TRIM37 and HUWE1 were detected by qRT-PCR and western blotting in OC cells. Cell proliferation was evaluated by CCK-8 assay and colony formation assay. Cell migration and invasion capabilities were examined by wound healing and transwell assay. Flow cytometry and western blotting were performed to measure cell apoptosis. Wnt/β-catenin pathway and the expression of c-Myc were identified by qRT-PCR and western blotting. The binding of TRIM37 and HUWE1 was predicted by STRING database and verified by co-immunoprecipitation. In addition, a xenograft mouse model was established to evaluate the effects of TRIM37 and HUWE1 on tumor growth and c-MYC expression in vivo. The present study revealed that TRIM37 expression was upregulated in OC tissues and cells. TRIM37 silencing inhibited OC cell migration and invasion, promoted OC cell apoptosis, and blocked Wnt/β-catenin signaling pathway, as well as suppressing c-MYC protein expression. Mechanistic studies suggested that TRIM37 binds to HUWE1. HUWE1 was upregulated in OC cells and TRIM37 promoted the c-MYC expression through targeting HUWE1. Animal experiments showed that TRIM37 silencing significantly repressed the tumor growth and c-MYC protein level, but HUWE1 overexpression reversed the effects of TRIM37 knockdown on mice with OC. Our findings revealed that TRIM37 accelerated the progression of OC and promoted c-MYC expression by binding to HUWE1, which provides therapeutic targets for OC treatment.

Effect of substrate stiffness on the mechanical properties of cervical cancer cells

Cervical cancer microenvironment is involved in the regulation of the behavior, morphology, and mechanical properties of the cervical cancer cells. Integrins expressed on the cell membrane combine with the factors of the microenvironment to determine cervical cancer cells' properties. The mechanical properties of integrin-extracellular matrix (ECM) interactions are important for the mechanotransduction of cervical cancer cells. However, the quantified study on the adhesion force and binding probabilities between collagen and integrins on cervical cancer cells grown on different stiffness substrates have not been reported. Polyacrylamide (PA) gel was used as substrate to mimic the mechanical microenvironment of cancer cells. ImageJ software was used to measure the perimeter and area of the cells. SiHa cells were stained with FITC phalloidine to observe the cytoskeleton. Atomic force microscopy (AFM) was used to measure the cell mechanical properties. The perimeters of SiHa cells grown on different stiffness substrates were 63.4 ± 1.3, 102.8 ± 4.0, and 152.6 ± 4.1 μm, for soft, intermediate, and stiff substrates, respectively. These areas were 277.2 ± 13.3, 428.9 ± 26.0, and 1166.0 ± 63.2 μm The changes in substrate stiffness can obviously regulate SiHa cells' mechanical properties, such as the Young's modulus. The adhesion force and binding probabilities of SiHa cells both increased with increasing substrate strength.

Dysregulated circular RNA and long non-coding RNA-Mediated regulatory competing endogenous RNA networks (ceRNETs) in ovarian and cervical cancers: A non-coding RNA-Mediated mechanism of chemotherapeutic resistance with new emerging clinical capacities

Cervical cancer (CC) and ovarian cancer (OC) are among the most common gynecological cancers with significant mortality in women, and their incidence is increasing. In addition to the prominent role of the malignant aspect of these cancers in cancer-related women deaths, chemotherapy drug resistance is a major factor that contributes to their mortality and presents a clinical obstacle. Although the exact mechanisms behind the chemoresistance in these cancers has not been revealed, accumulating evidence points to the dysregulation of non-coding RNAs (ncRNAs), particularly long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs), as key contributors. These ncRNAs perform the roles of regulators of signaling pathways linked to tumor formation and chemoresistance. Strong data from various recent studies have uncovered that the main mechanism of these ncRNAs in the induction of chemoresistance of CC and OC is done through a dysregulated miRNA sponge activity as competing endogenous RNA (ceRNA) in the competing endogenous RNA networks (ceRNETs), where a miRNA regulating a messenger RNA (mRNA) is trapped, thereby removing its inhibitory effect on the desired mRNA. Understanding these mechanisms is essential to enhancing treatment outcomes and managing the problem of drug resistance. This review provides a comprehensive overview of lncRNA- and circRNA-mediated ceRNETs as the core process of chemoresistance against the commonly used chemotherapeutics, including cisplatin, paclitaxel, oxaliplatin, carboplatin, and docetaxel in CC and OC. Furthermore, we highlight the clinical potential of these ncRNAs serving as diagnostic indicators of chemotherapy responses and therapeutic targets.

Estrogen promotes Epithelial ovarian cancer cells proliferation via down-regulating expression and activating phosphorylation of PTEN

Epithelial ovarian cancer (EOC) is the most common of cancer death among malignant tumors in women, its occurrence and development are strongly linked to estrogen. Having identified the phosphatase and tensin homologue (PTEN) is a potent tumor suppressor regulating cell proliferation, migration, and survival. Meanwhile, there is a correlation between PTEN protein expression and estrogen receptor expression in EOC. However, no study has amplified on the molecular regulatory mechanism and function between estrogen and PTEN in the development of EOC. In this research, we found that PTEN shows a low expression level in EOC tissues and estrogen decreased PTEN expression via the estrogen receptor 1 (ESR1) in EOC cells. Knockdown of PTEN enhanced the proliferation and migration level of EOC cells driven by estrogen. Moreover, PTEN was also phosphorylated by G protein-coupled receptor 30 (GPR30)-Protein kinase C (PKC) signaling pathway upon estrogen stimulation. Inhibiting the phosphorylation of PTEN weakened the proliferation and migration of estrogen induced-EOC cells estrogen and decreased the phosphorylation of Protein kinase B (AKT) and Mammalian target of rapamycin (mTOR). These results indicated that estrogen decreased PTEN expression level via the ESR1 genomic pathway and phosphorylated PTEN via the GPR30-PKC non-genomic pathway to activate the PI3K/AKT/mTOR signaling pathway, thereby determining the fate of EOC cells.

LncRNA SLC2A1-AS1 facilitates tumor progression by targeting miR-508-5p/ FOXO1 axis in ovarian cancer cells

Epithelial ovarian cancer (EOC) has high tumor drug resistance, recurrence rate, and risk of metastasis. To elucidate the role of the long non-coding RNA SLC2A1-AS1 in EOC. A cohort of 140 EOC patients was enrolled. SLC2A1-AS1 and related transcripts were quantified by RT-qPCR. A panel of OC cell lines was then employed, and the oncogenic functions of SLC2A1-AS1 were systematically interrogated through CCK-8 proliferation, Transwell migration/invasion, and western blot analysis. Dual-luciferase reporter and RIP assays were performed to confirm the targeting interactions. SLC2A1-AS1 expression was significantly increased in EOC tissues. High expression of SLC2A1-AS1 significantly increased the recurrence rate and mortality. Multifactor Cox suggested that high expression of SLC2A1-AS1 was an independent risk factor for overall survival (OS) and recurrence-free survival (RFS). In addition, SLC2A1-AS1 targeted miR-508-5p, which was generally down-regulated in EOC and was significantly negatively correlated with SLC2A1-AS1. Furthermore, miR-508-5p directly targeted FOXO1, a gene markedly up-regulated in EOC and inversely correlated with miR-508-5p levels. Depletion of SLC2A1-AS1 liberated miR-508-5p, which in turn binds FOXO1 mRNA, thereby suppressing proliferation, migration, and invasion of SKOV3 and OVCAR3 cells. This tumor-suppressive program could be largely weakened by miR-508-5p-inhibitor, underscoring the SLC2A1-AS1/miR-508-5p/FOXO1 axis as a critical driver of EOC aggressiveness. Consistent results were obtained in additional EOC cell lines A2780 and CAOV3, confirming the generalizability of the SLC2A1-AS1/miR-508-5p/FOXO1 axis across different cellular backgrounds. SLC2A1-AS1 may drive the progression and recurrence of EOC through the miR-508-5p/FOXO1 axis.

RHBDD1 promotes cervical cancer progression by activating the EGFR/PI3K/AKT signaling pathway

Although RHBDD1 has been linked to tumorigenesis, its specific role in cervical cancer (CC) remains elusive. This study investigated its oncogenic function and regulatory mechanism in CC cells. CC cell lines were transfected with either RHBDD1-targeting or overexpression constructs. A series of functional assays, including CCK-8, colony formation, flow cytometry, and Transwell assays, were conducted to assess proliferation, cell cycle progression, apoptosis, migration, and invasion. Pharmacological modulators targeting the EGFR/PI3K/AKT pathway-gefitinib (EGFR inhibitor, 10 μM), LY294002 (PI3K inhibitor, 20 μM), and SC79 (AKT activator, 10 μM)-were applied in combination with gene manipulation. Western blotting was performed to assess pathway activation and expression of EMT-related proteins, including Snail. Co-immunoprecipitation (Co-IP) assays were conducted to evaluate the physical association between RHBDD1 and EGFR. Additionally, a xenograft mouse model was used to evaluate tumorigenic potential in vivo. RHBDD1 knockdown in SiHa cells markedly impaired cell growth, invasion, and EMT, as evidenced by decreased Snail expression, whereas RHBDD1 overexpression in HeLa cells enhanced malignant phenotypes and upregulated Snail. Corresponding changes in EGFR/PI3K/AKT signaling were observed: pathway activity was elevated with RHBDD1 overexpression and suppressed upon silencing. Co-IP assays confirmed a physical association between RHBDD1 and EGFR, supporting receptor-level modulation. Notably, SC79 partially reversed the suppressive effects of RHBDD1 knockdown, whereas gefitinib and LY294002 mitigated RHBDD1-induced oncogenicity. These findings were corroborated by alterations in AKT phosphorylation levels. In vivo, RHBDD1 depletion significantly reduced tumor growth, EMT marker expression, and pathway activation. RHBDD1 facilitates CC progression by promoting proliferation, EMT, migration, invasion, and tumorigenesis through activation of the EGFR/PI3K/AKT pathway, likely via direct receptor-level interaction. Targeting this regulatory node may offer a promising therapeutic approach for CC.

Targeting the METTL3/YTHDF3/m6A/PGK1 axis to combat Choriocarcinoma progression

Choriocarcinoma (CC) is a highly aggressive trophoblastic tumor characterized by rapid progression and early metastasis. Glycolysis, a crucial metabolic pathway in cancer, is essential for tumor growth, yet its specific role in CC is not well understood. Phosphoglycerate kinase 1 (PGK1), an important enzyme in glycolysis, along with N6-methyladenosine (m6A) RNA methylation, a common modification of mRNA, are both associated with tumorigenesis. However, the role of m6A-modified PGK1 in CC has not been investigated. In this study, bioinformatics analysis has identified PGK1 as a key regulator in the progression of CC and glycolysis. PGK1 was found to be upregulated in CC cell lines BeWo and JEG-3 cells. Functional experiments indicated that PGK1 knockdown suppressed proliferation and glycolysis of tumor cells, and inhibited tumor growth in xenograft mouse models. Mechanistically, the m6A modification of PGK1 mRNA, mediated by methyltransferase-like 3 (METTL3), increased its stability, while YTH domain-containing family protein 3 (YTHDF3) promoted its translation in an m6A-dependent manner. Silencing PGK1 diminished tumor progression, while the overexpression of METTL3 or YTHDF3 partially mitigated these effects by restoring PGK1 expression and glycolytic function. Overall, the METTL3/YTHDF3 axis enhanced CC progression by stabilizing PGK1 mRNA through m6A modification. These results position PGK1 as a potential diagnostic biomarker and therapeutic target, indicating that targeting the METTL3/YTHDF3/m6A/PGK1 pathway may provide innovative strategies for the treatment of CC.

Norcantharidin inhibits the METTL16/MAT2A pathway to induce apoptosis and suppress tumor progression in ovarian cancer

This study aimed to investigate the antitumor effects of norcantharidin (NCTD) in ovarian cancer and to elucidate its molecular targets and mechanisms of action, with a focus on the methyltransferase-like protein 16 (METTL16)/methionine adenosyl transferase II alpha (MAT2A) signaling axis. Expression levels of METTL16 and MAT2A were analyzed in ovarian cancer tissues and cells using immunohistochemistry, quantitative real-time polymerase chain reaction, and western blotting. Human ovarian cancer cell lines (ES2 and SKOV3) were cultured and subjected to METTL16 overexpression via gene recombination. The cytotoxic effects of varying concentrations of NCTD were assessed using the Cell Counting Kit-8 assay. Apoptosis, proliferation, and migration were evaluated through flow cytometry and wound healing (scratch) assays. A subcutaneous xenograft model of human ovarian cancer was established in nude mice to assess in vivo antitumor efficacy. Enzyme-linked immunosorbent assay was used to quantify intracellular levels of S-adenosylmethionine (SAM). METTL16 and MAT2A gene expression levels were significantly elevated in ovarian cancer tissues and cell lines (p < 0.05). Treatment with NCTD at 10 μg/mL significantly inhibited proliferation and induced apoptosis in ES2 and SKOV3 cells. NCTD also suppressed cellular migration and angiogenic activity, with downregulation of related gene expression, effects that were attenuated by METTL16 overexpression (p < 0.05). In the xenograft model, NCTD administration significantly reduced tumor volume and downregulated expression of METTL16, MAT2A, protein phosphatase 2A (PP2A), and vascular endothelial growth factor (p < 0.05). NCTD exerts antineoplastic effects in ovarian cancer by reducing intracellular S-adenosylmethionine levels, promoting PP2A demethylation, and inhibiting the METTL16/MAT2A signaling pathway. These effects contribute to cell cycle arrest, suppressed proliferation, and enhanced apoptosis, supporting the therapeutic potential of NCTD in ovarian malignancies.

SIRT7 deficiency promoted cuproptosis-mediated mitochondrial dysfunction and inhibited malignant development of cervical cancer

To investigate the impact of SIRT7 on the development of cervical cancer and its relationship with cuproptosis in cervical cancer. HeLa and SiHa cells were transfected with lentiviruses for SIRT7 overexpression and knockdown. The effects of SIRT7 on cervical cancer cell proliferation, apoptosis, invasion, and migration were analyzed using CCK8, plate cloning, flow cytometry, Transwell assays, and scratch assays. To verify the relationship between SIRT7 and cuproptosis, we utilized cuproptosis inhibitors and activators. Immunofluorescence, transmission electron microscope, flow cytometry, ELISA, and Western blot were used to analyze copper ion content, mitochondrial ultrastructure, cellular reactive oxygen species, mitochondrial membrane potential, pyruvate levels, cell viability and the cuproptosis-related proteins. SIRT7 enhanced the proliferation, migration, and invasion of HeLa and SiHa cells, inhibited apoptosis, and promoted cervical cancer growth. Knocking down SIRT7 caused cuproptosis of HeLa and SiHa cells, characterized by increased Cu SIRT7 was recognized as an oncogene in cervical cancer, which boosted cervical cancer cell proliferation and invasion, lowered intracellular copper levels, and prevented cuproptosis. SIRT7 downregulation triggered cuproptosis, inhibiting tumor cell growth.

Exosomal miR16 induced by allyl isothiocyanate (AITC) inhibits tumor growth in cervical cancer via modulation of apoptotic and inflammatory pathways

The tumor micro-environment is a key determinant for promoting cancer cell growth and development with exosomal miRNAs emerging as key regulators of tumor growth and metastasis. miR16 is one well-established tumor suppressor miRNAs that induces apoptosis, while inhibiting angiogenesis and inflammation across various cancers. Herein, we investigated the role of exosomal miR16 in the cervical cancer microenvironment and its underlying molecular mechanisms. We treated human cervical cancer HeLa cells with Allyl Isothiocyanate (AITC) and observed the impact of miR16-enriched exosomes on human fibrosarcoma HT1080 cells. We found a significant increase of miR16 expression in AITC-treated HeLa cells and purified exosomes. When the exosomes were cultured with fibroblasts, miR16 expression was increased in fibroblast cells. Treatment with AITC-exposed HeLa exosomes induced increased Bax/Bcl2 ratio and downregulated PCNA, HIF-1α, SDF-1α, IL-6, and p22phox expression in fibroblasts. Remarkably, the knockdown of miR16 in fibroblasts inhibited the AITC-induced increase in the Bax/Bcl2 ratio and restored VEGF, PCNA, HIF-1α, SDF-1α, IL-6, and p22phox expression. In sum, our findings demonstrate the potential of AITC-mediated exosomal miR16 enrichment as an effective approach to inhibit cancer growth and development, and reveal a new potential for cancer management and therapy.

CircSOS2 promotes cervical squamous cell carcinoma by regulation of proliferation, cell cycle, apoptosis, migration, invasion, and glycolysis by targeting miR-543/FNDC3B axis

Cervical squamous cell carcinoma (SCC) is a common subtype of cervical cancer. Circular RNAs (circRNAs) have been demonstrated as vital regulators in gene regulation and malignant tumor progression. Therefore, the precise role of circular RNA salt overly-sensitive 2 (circSOS2) was investigated in SCC. The relative expression levels of circSOS2, microRNA-543 (miR-543), and Fibronectin type III domain containing 3B (FNDC3B) were determined by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot assays. The correlation between percent survival times of SCC patients and circSOS2 level was presented by Kaplan-Meier Plotter analysis. The cell proliferation was measured by MTT and colony-forming assays. Flow cytometry assay was used to assess apoptosis and cell cycle distribution. The migration and invasion were measured by transwell assay. The glycolysis was analyzed by extracellular acidification rate (ECAR) assay, Glucose Assay Kit, and Lactate Assay Kit. The interaction relationship between miR-543 and circSOS2 or FNDC3B was analyzed by dual-luciferase reporter, RNA immunoprecipitation (RIP), and RNA pull-down assays. A xenograft experiment was established to clarify the functional role of circSOS2 inhibition in viv. CircSOS2 was highly expressed in SCC tissues and cells; besides, its expression level was closely associated with poor prognosis. Loss-of-functional experiments revealed that suppression of circSOS2 repressed proliferation, cell cycle process, migration, invasion, and glycolysis while induced apoptosis in SCC cells, which was overturned by inhibition of miR-543. In addition, miR-543 was downregulated and negatively correlated with circSOS2 expression in SCC tissues. We also found that overexpression of miR-543 impeded proliferation, cell cycle process, migration, invasion, and glycolysis while induced apoptosis in SCC cells by targeting FNDC3B. The silencing of circSOS2 impeded tumorigenesis in vivo. CircSOS2 conferred an oncogenic function in SCC by regulation of proliferation, cell cycle, apoptosis, migration, invasion, and glycolysis of SCC cells, which was contributed to its interactions with miR-543 and FNDC3B.

APLN promotes the proliferation, migration, and glycolysis of cervical cancer through the PI3K/AKT/mTOR pathway

Apelin (APLN) is an endogenous ligand of the G protein-coupled receptor APJ (APLNR). APLN has been implicated in the development of multiple tumours. Herein, we determined the effect of APLN on the biological behaviour and underlying mechanisms of cervical cancer. The expression and survival curves of APLN were determined using Gene Expression Profiling Interactive Analysis. The cellular functions of APLN were detected using CCK-8, clone formation, EdU, Transwell assays, flow cytometry, and seahorse metabolic analysis. The underlying mechanisms were elucidated using gene set enrichment analysis and Western blotting. APLN was upregulated in the samples of patients with cervical cancer and is associated with poor prognosis. APLN knockdown decreased the proliferation, migration, and glycolysis of cervical cancer cells. The opposite results were observed when APLN was overexpressed. Mechanistically, we determined that APLN was critical for activating the PI3K/AKT/mTOR pathway via APLNR. APLN receptor inhibitor ML221 reversed the effect of APLN overexpression on cervical cancer cells. Treatment with LY294002, the PI3K inhibitor, drastically reversed the oncological behaviour of APLN-overexpressing C-33A cells. APLN promoted the proliferation, migration, and glycolysis of cervical cancer cells via the PI3K/AKT/mTOR pathway.

Calycosin inhibits viability, induces apoptosis, and suppresses invasion of cervical cancer cells by upregulating tumor suppressor miR-375

Calycosin, a functional phytoestrogen isoflavone isolated from Radix astragali, has been shown to possess multiple pharmacological properties including anti-cancer activity. However, up to now, the anti-cancer effect and the related mechanism of calycosin on cervical cancer (CC) cells have not been explored. It has been demonstrated that tumor suppressor miR-375 was downregulated in CC and calycosin upregulated miR-375 expression in cerebral ischemia/reperfusion. Thus we supposed that calycosin exerted anti-cancer effect by upregulating miR-375 expression in CC cells. Effects of calycosin or combined with miR-375 on cell viability and lactate dehydrogenase (LDH) release were detected by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetra zoliumromide (MTT) and LDH release assay. Apoptosis, caspase-3 activity, and cell invasion were determined by flow cytometry, caspase-3 activity assay, and Transwell assay, respectively. miR-375 expression was detected by quantitative real-time PCR (qRT-PCR). Our results showed that Calycosin dose-dependently inhibited cell viability and increased LDH release in CC cells, suggesting the cytotoxic effect of calycosin on CC cells. Calycosin enhanced the apoptotic rate and caspase-3 activity and decreased the number of invaded cells in CC cells. In addition, we found that miR-375 expression was decreased in CC cells but was upregulated in response to calycosin. Mechanistically, knockdown of miR-375 significantly reversed the anti-cancer effect of calycosin on CC cells. In conclusion, calycosin inhibited viability, induced apoptosis, and suppressed invasion of CC cells by upregulating tumor suppressor miR-375.

The roles of microRNA in human cervical cancer

Although a potentially preventable disease, cervical cancer (CC) is the second most commonly diagnosed gynaecological cancer with at least 530,000 new cases annually, and the prognosis with CC is still poor. Studies suggest that aberrant expression of microRNA (miRNA) contributes to the progression of CC. As a group of small non-coding RNA with 18-25 nucleotides, miRNA regulate about one-third of all human genes. They function by repressing translation or inducing mRNA cleavage or degradation, including genes involved in diverse and important cellular processes, including cell cycling, proliferation, differentiation, and apoptosis. Results showed that misexpression of miRNA is closely related to the onset and progression of CC. This review will provide an overview of the function of miRNA in CC and the mechanisms involved in cervical carcinogenesis.

Long non-coding RNA GAS5 regulates the growth and metastasis of human cervical cancer cells via induction of apoptosis and cell cycle arrest

Studies have proved the role of GAS5 in the development of different cancers. This study was undertaken to investigate the role and explore therapeutic implications of GAS5 in human cervical cancer. The results showed that GAS5 was significantly (p < 0.05) downregulated in human cervical cancer tissues. The results also showed that cervical cancer progresses with the suppression of GAS5 expression levels. Additionally, the expression of GAS5 was also significantly (p < 0.05) downregulated in human cervical cancer cell lines. Nonetheless, overexpression of GAS5 caused a remarkable decrease in the proliferation of C33A and HeLa cervical cancer cells. The decrease in the proliferation rate was attributed to the induction of apoptosis of C33A and HeLa cells which was accompanied with upregulation of Bax and suppression of Bcl-2. Additionally, GAS5 overexpression also promoted the arrest of C33A and HeLa cells at the G2/M check point of cell cycle via suppression of cyclin B1 and CDK1 expression. The transwell assays showed that GAS5 overexpression significantly (p < 0.05) inhibited the migration and invasion of the C33A and HeLa cervical cancer cells. The bioinformatics analysis as well as the dual luciferase assay showed GAS5 acts as a target of miR-135a. Interestingly, the expression of miR-135a was upregulated in the human cervical cancer cells and its suppression exerted growth inhibitory effects on the C33A and HeLa cells. However, silencing of GAS5 could nullify the effects of miR-135a suppression on the proliferation of C33A and HeLa cells. Taken together, the results of this study point towards the therapeutic implications of GAS5 in the treatment of cervical cancer.

FKBP38 suppresses endometrial cancer cell proliferation and metastasis by inhibiting the mTOR pathway

Endometrial cancer (EC) is a common gynecological malignancy, and advanced-stage or recurrent EC is associated with a high mortality rate owing to the ineffectiveness of currently available treatments. FK506-binding protein 38 (FKBP38) is a member of the immunophilin family and inhibits melanoma and breast cancer cell metastasis. However, the functions of FKBP38 and its potential mechanism in EC remain unclear. Herein, we analyzed the expression levels of FKBP38 in EC cells and found that the FKBP38 expression was high in Ishikawa cells, and low in AN3CA cells, traditionally considered a low grade and a high grade cell line, respectively, in pathology classification. Moreover, FKBP38 inhibited cell proliferation, migration and invasion in EC cells, FKBP38 knockdown significantly promoted tumor growth of Ishikawa cells in a subcutaneous xenograft model and increased the number of lung metastases of Hec-1-A cells in a metastatic mouse model. Furthermore, FKBP38 suppressed several target proteins of epithelial-to-mesenchymal transition (EMT) and reduced the phosphorylation of ribosomal S6 protein (S6), eukaryotic initiation factor 4E-binding protein 1 (4EBP-1), indicating the potent inhibition of the mammalian target of rapamycin (mTOR) pathway. Meanwhile, the inhibition of mTOR neutralized the elevation of EC cell proliferation, migration and invasion after FKBP38 knockdown. In summary, FKBP38 would exert a tumor-suppressing role by modulating the mTOR pathway. Our results indicate that FKBP38 may be considered as a factor of EC metastasis and a new target for EC therapeutic intervention.

Lactamase β reprograms lipid metabolism to inhibit the progression of endometrial cancer through attenuating MDM2-mediated p53 ubiquitination and degradation

Lactamase β (LACTB) inhibits the metastasis and progression of multiple malignant tumors. However, little is known about its role in endometrial cancer (EC). Our study aimed to investigate the function and potential molecular mechanism of LACTB in modulating EC progression. LACTB expression was measured via immunohistochemistry staining, Western blot and qRT-PCR. The role of LACTB in EC was investigated both in vivo and in vitro by employing xenograft mice models and using colony formation, EdU, and Transwell assays, along with flow cytometric analysis. In addition, to assess LACTB function on lipid metabolism, lipid droplets in EC cells were labeled with Nile red. Western blot, immunofluorescence staining, co-immunoprecipitation, ubiquitination assay, and cycloheximide chase assay and rescue experiments were performed to confirm the interaction between LACTB, p53, and MDM2 in EC. LACTB expression was downregulated in EC. LACTB inhibited the malignant phenotypes and reprogramed lipid metabolism in EC cells. Moreover, LACTB significantly upregulated p53 by attenuating the MDM2-mediated ubiquitination and degradation of p53. Besides, LACTB silencing facilitated the malignant phenotypes and reprogramed lipid metabolism in EC cells; this was reversed with p53 overexpression. LACTB knockdown facilitated EC progression via downregulating p53 in vivo. LACTB repressed EC cell proliferation and metastasis, and reprogramed lipid metabolism via attenuating the MDM2-mediated ubiquitination and degradation of p53.

SMC4 knockdown inhibits malignant biological behaviors of endometrial cancer cells by regulation of FoxO1 activity

Structural maintenance of chromosomes 4 (SMC4) has an important role in chromosome condensation and segregation, which is involved in regulating multiple tumor development. However, the role of SMC4 in endometrial cancer is uncertain. The expression and prognostic value of SMC4 were predicted by UALCAN, Gene Expression Omnibus (GEO), The Human Protein Atlas and Kaplan Meier plotter tools. SMC4-related genes were analyzed by LinkedOmics, Gene Ontology (GO) annotations, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Forkhead box protein O1 (FoxO1) activity was suppressed by AS1842856 (AS). SMC4, Ki67, B-cell lymphoma-2(Bcl-2), Bcl-2 associated X protein (Bax), FoxO1, phosphorylated FoxO1 (p-FoxO1), and p27 protein levels were detected by Western blotting. Cell proliferation was detected using Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) analyses. Cell apoptosis was measured using TUNEL analysis. SMC4 abundance was increased in endometrial cancer, and predicted a worse overall survival. SMC4 knockdown repressed proliferative ability of endometrial cancer cells and promoted cell apoptosis. SMC4 knockdown promoted FoxO1 transactivation by decreasing its phosphorylated level. Addition of AS inhibited FoxO1 activity by increasing the phosphorylated level of FoxO1. The inhibition of FoxO1 activity reversed the effect of SMC4 silencing on cell proliferation and apoptosis. In conclusion, SMC4 silencing restrained cell proliferation and facilitated apoptosis in endometrial cancer via regulating FoxO1 activity.