Journal of Molecular Histology

TMEM130 promoter hypermethylation predicts tumor cell migration in cervical cancer

Cervical cancer represents a prevalent malignancy affecting women globally, DNA methylation serves as a crucial epigenetic modification influencing disease prognosis through the regulation of gene expression. The aim of this study is to elucidate the potential impact of TMEM130 promoter methylation on its transcriptional activity and to explore the functional role of TMEM130 in the migration of cervical cancer cells. Clinical data and gene expression profiles from The Cancer Genome Atlas (TCGA) were analyzed to assess the correlation between TMEM130 expression and patient overall survival. Transcriptional levels of TMEM130 in cervical cancer cell lines and clinical specimens were quantified via qRT-PCR, revealing significant downregulation relative to normal controls, consistent with the TCGA findings. Reduced TMEM130 expression was associated with poorer overall survival. Bisulfite sequencing PCR (BSP) confirmed hypermethylation of the TMEM130 promoter in tumor tissues and treatment with a methyltransferase inhibitor restored TMEM130 expression. Functional assays, including Transwell migration and Western blot analyses following TMEM130 plasmid transfection, demonstrated that TMEM130 overexpression suppressed migratory capabilities in cervical cancer cells. These findings suggested that in cervical cancer TMEM130 might act as a tumor suppressor, with promoter hypermethylation contributing to its downregulation, promising its potential as a biomarker for disease progression.

MUL1 suppresses cervical cancer progression by targeting FUNDC1 for ubiquitination and inhibiting DRP1-dependent mitophagy

Cervical cancer is the fourth most common cancer in women worldwide. Mitochondrial E3 ubiquitin ligase 1 (MUL1) plays a crucial role in cancer processes, yet its role in cervical cancer remains unclear. Here, we observed that MUL1 mRNA and protein levels were reduced in cervical cancer tissues and cells using qRT-PCR and Western blot assays. Patients with low MUL1 expression exhibited poor survival. CCK-8, EdU, Transwell and flow cytometry analysis demonstrated that MUL1 overexpression inhibited cervical cancer cell proliferation and migration and promoted cell apoptosis, while MUL1 knockdown had opposite effects. Interestingly, inhibition of mitophagy induced by Midivi-1 attenuated the effects of MUL1 knockdown on cell proliferation, migration, and apoptosis. Mechanistically, co-immunoprecipitation and ubiquitination assays demonstrated that MUL1 decreased FUN14 domain containing 1 (FUNDC1) protein stability by promoting its ubiquitination. FUNDC1 overexpression promoted dynamin-related protein 1 (DRP1) expression and promoted mitophagy in cervical cancer cells, whereas DRP1 knockdown reversed these changes. Notably, FUNDC1 knockdown weakened the promoting effects of MUL1 knockdown on mitophagy. FUNDC1 overexpression rescued the inhibitory effects on proliferation, migratory capacity and the promoting effect on apoptosis. In vivo, MUL1 overexpression inhibited tumor growth in a xenograft mouse model. These findings suggested MUL1 suppressed cervical cancer progression by targeting the FUNDC1/DRP1 axis and inhibiting mitophagy, highlighting its potential as a therapeutic target.

LOXL1-AS1 suppresses ferroptosis in cervical cancer through m6A-dependent regulation of TFRC

Transferrin receptor (TFRC) is essential for iron uptake and may regulate ferroptosis, a form of iron-dependent cell death implicated in cervical cancer (CC). Bioinformatic analyses (GEPIA, UALCAN, SRAMP, starBase) were combined with in vitro and in vivo experiments. CC cell lines were transfected with shRNAs or overexpression plasmids targeting TFRC and LOXL1-AS1. Proliferation was assessed by colony formation, EdU staining, and Ki-67 immunostaining. Ferroptosis was evaluated by measuring malondialdehyde (MDA), lipid ROS, Fe

Role of indoleamine 2,3-Dioxygenase 1 in modulating the malignant biological behavior of cervical cancer cells through the Tryptophan-Kynurenine pathway

The aim of this study is to evaluate the influence of altered expression of indoleamine 2,3-dioxygenase 1 (IDO1) on key enzymes and metabolites within the tryptophan-kynurenine pathway, as well as to assess the resulting effects on the malignant biological behavior of cervical cancer cells, including proliferation, invasion and migration. IDO1 expression was assessed in cervical cancer and normal cervical tissues via immunohistochemistry and western blot (WB) analysis. Four human cervical cancer cell lines (HeLa, C33A, SiHa, CaSki) were cultured, and HeLa and SiHa cells-characterized by elevated IDO1 expression-were selected for further experimentation. IDO1 was silenced using RNA interference techniques. Quantitative real-time polymerase chain reaction and WB were used to detect the messenger RNA and protein expression levels of IDO2, tryptophan 2,3-dioxygenase (TDO), kynurenine 3-monooxygenase (KMO), and aryl hydrocarbon receptor (AhR). Targeted metabolomics analysis based on liquid chromatography-tandem mass spectrometry was conducted to quantify tryptophan, N-formylkynurenine (NFK), and L-kynurenine (L-KYN) in cell culture supernatants. Cell proliferation, apoptosis, and migratory capacity were evaluated via flow cytometry and scratch assays. IDO1 expression was significantly elevated in cervical cancer tissues compared to normal cervical tissues, with the highest levels observed in samples positive for human papillomavirus types 16 or 18. Silencing of IDO1 in cervical cancer cells resulted in downregulation of IDO2, TDO, KMO, and AHR expression. Correspondingly, reduced metabolic activity in the tryptophan-kynurenine pathway was observed, evidenced by decreased levels of NFK and L-KYN. These molecular alterations were associated with G1 phase cell cycle arrest, diminished cellular proliferation, migration and invasion, and increased apoptotic activity. IDO1 mediates the malignant biological behavior of cervical cancer cells through the tryptophan-kynurenine pathway, suggesting its potential as a therapeutic target in cervical malignancies.

Evaluating TGF-β1 gene expression and promoter polymorphism in cervical cancer progression

This study aims to investigate the TGF-β1 gene, which has significant prognostic value for early detection and diagnosis of cervical cancer, as well as TGF-β1 gene mRNA and protein expression and the association of promoter region (-509 C>T) polymorphisms with cervical cancer (CC) development. Transcriptome analysis, immunohistochemistry, and RT-PCR were conducted to determine the gene expression of TGF-β1. The PCR-SSCP and Sanger sequencing methods were employed to test and validate the TGF-β1 -509C>T promoter polymorphism in cervical squamous cell carcinoma in comparison to control samples. TGF-β1 is a cytokine that plays a role in tumorigenesis as well as physiological and pathological processes. It appeared as one of the most over-expressed genes identified through the clariom D transcriptome microarray, which describes its role in cancer progression. The results showed a significant TGF-β1 upregulation in CC compared to normal cervical tissue was confirmed using immunohistochemistry and real-time PCR. The levels of TGF-β1 were also determined using a receiver operating characteristic (ROC) curve to distinguish diseased from normal individuals. TGF-β1 ROC showed good selectivity in distinguishing malignant CC from non-malignant cervical tissues. The -509 C>T promoter polymorphism in the TGF-β1 gene is found to be significantly more common in the disease group, and in-silico analysis (using the AliBaba2.0 gene regulation tool) confirms its correlation to the loss of myogenin transcription factor binding site, may resulting in TGF-β1 overexpression.

RING box protein-1 promotes the metastasis of cervical cancer through regulating matrix metalloproteinases via PI3K/AKT signaling pathway

Cervical cancer (CC) remains a leading cause of cancer mortality amongst females worldwide. Some of the CC patients may experience early metastases of the primary tumor, however the underlying mechanism remains unclear. Aberrant expression of RING box protein-1 (RBX1), a subunit in the E3 ubiquitin ligase family, has been reported in several cancer types. Nevertheless, little is known regarding the role of RBX1 in the metastasis of CC patients. In this study, we examined the expression of RBX1 from 90 biopsies of CC patients, and found a significantly increased expression of RBX1 in the tumor tissues compared to the normal tissues. Notably, the abundance of RBX1 in the CC patients with metastasis was higher than their counterparts without metastasis, suggesting that RBX1 may play a significant role in the modulation of CC metastasis. Furthermore, by using Hela cells as a model of CC in vitro, we demonstrated that ectopic over-expression of RBX1 could significantly promote the migration and invasion of Hela cells, whereas knockdown of RBX1 could remarkably suppress the migration and invasion of Hela cells. Mechanistically, the regulatory effect of RBX1 on cell metastasis was associated with changes in matrix metalloproteinases (MMP3 and MMP9) and altered activity of PI3K/AKT signaling. In conclusion, this study highlighted RBX1 as a novel target that can promote the metastasis of Hela cells in vitro, which may contribute to the development of alternative therapeutic options for CC patients.

Roles of epidural block in combination with general anesthesia in stress response and immune function of patients after surgery for cervical cancer

We aimed to explore the roles of epidural block in combination with general anesthesia in the stress response and immune function of patients after surgery for cervical cancer. A total of 108 patients undergoing radical surgery of cervical cancer were randomly assigned into a general anesthesia combined with epidural block (observation) group and a general anesthesia (control) group. Peripheral blood was collected before anesthesia (t0), during anesthesia maintenance, as well as 10 min, 1 d, 2 d and 7 d after surgery. The levels of cytokines interferon-γ (IFN-γ), interleukin-4 (IL-4) and transforming growth factor-β1 (TGF-β1) were detected by ELISA, and IFN-γ/IL-4 ratio was calculated. Compared with the control group, the observation group had significantly lower levels of GH, PRL and Cor, proportions of Th2 and Treg cells, and levels of IL-4 and TGF-β1 during anesthesia maintenance and at each time point after surgery (P < 0.05), but higher proportion of Th1 cells, Th1/Th2 cell ratio, IFN-γ level and IFN-γ/IL-4 ratio (P < 0.05). General anesthesia in combination with epidural block can work better in mitigating the stress response and protecting the immune function of patients after cervical cancer surgery.

Germ cell-specific gene 2 accelerates cell cycle in epithelial ovarian cancer by inhibiting GSK3α-p27 cascade

AbstractEpithelial ovarian cancer (EOC) is one of the most common malignant gynecological tumors with rapid growth potential and poor prognosis, however, the molecular mechanism underlying its outgrowth remained elusive. Germ cell-specific gene 2 (GSG2) was previously reported to be highly expressed in ovarian cancer and was essential for the growth of EOC. In this study, GSG2-knockdown cells and GSG2-overexpress cells were established through lentivirus-mediated transfection with Human ovarian cancer cells HO8910 and SKOV3. Knockdown of GSG2 inhibited cell proliferation and induced G2/M phase arrest in EOC. Interestingly, the expression of p27, a well-known regulator of the cell cycle showed a most significant increase after GSG2 knockdown. Further phosphorylation-protein array demonstrated the phosphorylation of GSK3αSer21 decreased in GSG2-knockdown cells to the most extent. Notably, inhibiting GSK3α activity effectively rescued GSG2 knockdown’s suppression on cell cycle as well as p27 expression in EOC. Our study substantiates that GSG2 is able to phosphorylate GSK3α at Ser21 and then leads to the reduction of p27 expression, resulting in cell cycle acceleration and cell proliferation promotion. Thus, GSG2 may have the potential to become a promising target in EOC.

The expression and mechanism of action of ADAMTS18 in endometrial cancer

We aimed to measure ADAMTS18 expression in endometrial carcinoma (EC), atypical hyperplasia (AH), and normal endometrium, and determine its biological role in EC. Retrospectively, we analyzed clinicopathological data of the following groups: EC group (n = 64, endometrioid adenocarcinoma), AH group (n = 55), and control group (CON, n = 64, normal). ADAMTS18 expression was detected via immunohistochemical staining/immunofluorescence assay. Ishikawa EC cells were used in the following groups: ADAMTS18 group (overexpression plasmid), CON group (untreated), and NC group (null plasmid). The effects of ADAMTS18 on cell proliferation (CCK-8), migration/invasion (Transwell), and apoptosis (TUNEL) were assessed. ADAMTS18 expression was the lowest in the EC group and the highest in the CON group (P < 0.05). In Ishikawa cells, compared to the NC/CON groups, ADAMTS18 overexpression significantly decreased cell proliferation (after 72 h and 96 h), migration, and invasion, and enhanced cell apoptosis (all P < 0.05). Low ADAMTS18 expression was correlated with higher FIGO stage (≥ III) and larger tumor diameter (≥ 2 cm) in EC. ADAMTS18 downregulation was correlated with the poor prognosis of EC and suppressed tumor proliferation/invasion in vitro. ADAMTS18 overexpression modulated the behavior of EC cells by inhibiting their proliferation, invasion, and migration, and promoting their apoptosis. Functioning as a tumor suppressor, ADAMTS18 is a potential therapeutic target in EC.

CISD2 transcriptional activated by transcription factor E2F7 promotes the malignant progression of cervical cancer

Cervical cancer (CC) is the second most common type of cancer in women, and presents a serious threat to public health. We aimed to investigate the regulatory impacts of CDGSH iron-sulfur domain-containing protein 2 (CISD2) in CC and to discuss its relationship with E2F transcription factor 7 (E2F7). With the employment of real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and western blot, the expression of CISD2 and E2F7 in SiHa cells before or after transfection was estimated. Cell counting kit-8 (CCK-8) assay, Terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay, wound healing and transwell were used to detect the proliferation, apoptosis, migration and invasion of SiHa cells. The activity of CISD2 was detected using luciferase report assay and chromatin immunoprecipitation (ChIP) assay was used to confirm the binding of E2F7 and CISD2 promoter. The contents of proliferation- and apoptosis-related proteins were detected using western blot. Results revealed that CISD2 expression was greatly enhanced in CC cell lines. CISD2 depletion inhibited the proliferation, migration and invasion of SiHa cells but promoted the cell apoptosis. It was also found that E2F7 was remarkably elevated in SiHa cells. According to JASPAR database, the binding sites of E2F7 and CISD2 were predicted and ChIP confirmed the binding of E2F7 and CISD2 promoter. Results obtained from luciferase report assay indicated that E2F7 overexpression increased the activity of CISD2 promoter region. Furthermore, further functional experiments demonstrated that the impacts of E2F7 interference on the proliferation, migration, invasion and apoptosis of SiHa cells were reversed by CISD2 overexpression. In summary, CISD2 silence could alleviate the malignant progression of CC and could be transcribed by E2F7.

E2F1-Dependent CDCA5 overexpression drives cervical cancer progression and correlates with poor prognosis

Cervical cancer (CC) remains a leading cause of cancer-related mortality in women worldwide, highlighting the urgent need for novel therapeutic strategies. This study investigates the molecular mechanisms and clinical significance of Cell Division Cycle Associated 5 (CDCA5) in cervical cancer progression. We performed comprehensive analyses of CDCA5 expression in cervical cancer and normal tissues, correlating expression levels with clinicopathological features and patient outcomes. Functional studies using CC cell lines (SiHa, HeLa, and CaSki) examined the effects of CDCA5 manipulation on tumor cell behavior. We identified E2F1 as a key transcriptional regulator of CDCA5 and validated our findings using in vivo xenograft models. CDCA5 was significantly upregulated in CC tissues and correlated with advanced disease stages and poor survival outcomes. Mechanistically, CDCA5 depletion in SiHa and HeLa cells suppressed proliferation, migration, and invasion, while its overexpression in CaSki cells enhanced these malignant properties. We identified E2F1 as a transcriptional activator of CDCA5. Importantly, CDCA5 knockdown significantly inhibited tumor growth in nude mouse models. Our findings establish CDCA5 as a critical E2F1-regulated oncogenic factor in cervical cancer progression. The strong correlation between CDCA5 expression and poor clinical outcomes suggests its potential as both a prognostic biomarker and therapeutic target in cervical cancer treatment.

ELK1 activated-long noncoding RNA LBX2-AS1 aggravates the progression of ovarian cancer through targeting miR-4784/KDM5C axis

As one of the most common cancers in female, ovarian cancer (OC) has become a serious public burden now. Mounting researches have indicated long noncoding RNAs (lncRNAs) can affect many biological processes including cancer development. LncRNA LBX2-AS1 was identified to be an oncogene in some cancers, but the role of LBX2-AS1 in OC remains to be elucidated. Bioinformatics analysis and experiments including ChIP, RT-qPCR, RIP, luciferase reporter, western blot and CCK-8 were performed to explore the role of LBX2-AS1 in OC. LBX2-AS1 expression was markedly increased in OC tissues and cell lines. Functionally, LBX2-AS1 silencing inhibited cell proliferation, migration and stemness but facilitated cell apoptosis in OC. Moreover, depletion of LBX2-AS1 suppressed tumor growth of OC in vivo. Mechanically, LBX2-AS1 was activated by transcriptional factor ELK1. ELK1 enhanced the expression of LBX2-AS1 in OC cells. In addition, miR-4784 was confirmed to be sponged by LBX2-AS1. There was a negative expression correlation between LBX2-AS1 and miR-4784 in OC tissues. Subsequently, KDM5C was identified to be a direct target of miR-4784 in OC cells. KDM5C was negatively regulated by miR-4784 and positively regulated by LBX2-AS1 in terms of expression level. Upregulation of KDM5C reversed the inhibitory effect of LBX2-AS1 depletion on the progression of OC. This study proved that ELK1 activated-LBX2-AS1 aggravated the progression of OC by targeting the miR-4784/KDM5C axis, suggesting that LBX2-AS2 may be a promising diagnostic biomarker of OC.

Circular RNA RBM33 contributes to cervical cancer progression via modulation of the miR-758-3p/PUM2 axis

Cervical cancer (CC) is a gynecological malignant tumor. Circular RNA (hsa_circ_0001772) (circRBM33) is implicated in the tumorigenesis of cancers. Nevertheless, the role of circRBM33 in CC is indistinct. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to evaluate the levels of circRBM33, miR-758-3p, and pumilio RNA binding family member 2 (PUM2) mRNA in tissue samples and cells. Cell proliferation, apoptosis, migration, invasion, and glycolysis were assessed using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, flow cytometry assay, transwell assay, or special commercial kits. Relative protein levels were examined via western blotting. The targeting relationship between circRBM33 or PUM2 and miR-758-3p was verified via dual-luciferase reporter or RNA pull-down assays. The role of circRBM33 was confirmed via tumor formation experiments. CircRPPH1 and PUM2 were upregulated while miR-758-3p was downregulated in CC tissues and cells. Functionally, circRBM33 knockdown constrained tumor growth in vivo and cured CC cell proliferation, migration, invasion, glycolysis, and fostered CC cell apoptosis in in vitro. Mechanistically, circRBM33 sponged miR-758-3p to modulate PUM2 expression. MiR-758-3p inhibitor neutralized circRBM33 silencing-mediated effects on the malignant behaviors of CC cells. PUM2 elevation overturned the suppressive influence of miR-758-3p upregulation on the malignant behaviors of CC cells. CircRBM33 fostered CC advancement via absorbing miR-758-3p and upregulating PUM2, indicating that circRBM33 was a possible target for CC treatment.

Long non-coding RNA TDRG1 promotes hypoxia-induced glycolysis by targeting the miR-214-5p/SEMA4C axis in cervical cancer cells

Long non-coding RNA (lncRNA) has been demonstrated as vital regulator in human cancer. However, the precise role of lnc-TDRG1 in cervical cancer (CC) remains unclear, so this study was aimed to clarify the role and underlying molecular mechanism of lnc-TDRG1 in CC. The real-time quantitative polymerase chain reaction (RT-qPCR) was conducted to assess the expression levels of lnc-TDRG1, miR-214-5p and Semaphorin 4C (SEMA4C). Under hypoxia condition, the biological behaviors of CC cell, including invasion and glycolysis were determined by transwell assay and Glucose Assay Kit and Lactate Assay Kit, respectively. The Western blot assay was employed to test the expression level of SEMA4C and hexokinase 2 (HK2) expression. The interaction relationship between miR-214-5p and lnc-TDRG1 or SEMA4C was analyzed bioinformatics database and confirmed by dual-luciferase reporter assay, respectively. A xenograft experiment in nude mice was established to clarify the functional role of lnc-TDRG1 in vivo. We found Lnc-TDRG1 was highly expressed in CC tissues and cells and it was upregulated in response to hypoxia. Loss-of-functional experiment suggested that knockdown of lnc-TDRG1 impede invasion, hypoxia-induced glycolysis in vitro and tumor growth in vivo, which was abolished by knockdown of miR-214-5p or overexpression of SEMA4C. Moreover, we confirmed that miR-214-5p specifically bound to SEMA4C and negatively correlated with SEMA4C expression. Collectively, lnc-TDRG1 regulated SEMA4C expression by sponging miR-214-5p in CC. Collectively, mechanistically, lnc-TDRG1 could act as a sponge of miR-214-5p to regulate the expression of SEMA4C, and further regulate invasion and hypoxia-glycolysis in CC cells.

LncRNA expression signature correlated with cuproptosis: prognostic implications and insights into immune microenvironment in ovarian cancer

Ovarian cancer (OC) presents a formidable challenge in terms of early detection due to its subtle symptoms, often leading to diagnosis at advanced stages of the disease. Despite therapeutic advancements, survival rates exhibit limited improvement. This study delves into long non-coding RNAs (lncRNAs) associated with cuproptosis, aiming to anticipate the prognosis of OC and assess its immune status. The findings of this study provide a framework for enhancing the treatment of OC. RNA-seq and clinicopathological data of 379 TCGA-OC samples were retrieved from UCSC Xena. After extracting mRNAs/lncRNAs and screening CRLs via Pearson's analysis, a prognostic model was built and validated. Subgroup, PCA, GO/KEGG, immune infiltration, TMB, and drug sensitivity analyzes were conducted for evaluation. A prognostic ensemble, consisting of eight lncRNAs namely AC104820.2, EPB41L4A-AS1, LINC00996, RP11-110I1.6, RP11-367G6.3, RP11-443B7.3, RP11-4O1.2, and RP11-76E17.3, was effectively formulated. Independent prognostic factors for OC were identified through Cox analysis, with age and risk score emerging as noteworthy contributors. The prognostic signature demonstrated robust efficacy in the anticipation of 1-year, 3-year, and 5-year overall survival rates. Notably, individuals with low-risk OC manifested distinctive tumor immune microenvironments and exhibited an elevated tumor mutational burden. Furthermore, this low-risk cohort displayed heightened responsiveness to diverse therapeutic agents in comparison to their high-risk counterparts. Collectively, the comprehensive analysis of cuproptosis-associated lncRNAs serves a dual purpose in prognostication and elucidation of the immune microenvironment and therapeutic responsiveness in OC.

Endometrioid tubal intraepithelial neoplasia (E-TIN): case report &amp; literature review

An endometrioid carcinogenic pathway of the fallopian tube with possible potential precursors including type II SCOUTs (secretory cell outgrowths) and E-TIN (endometrioid tubal intraepithelial neoplasia) has been recently documented. We report an incidental focus of E-TIN identified in a hysterectomy specimen for Grade 1 endometrioid type endometrial carcinoma. The lesion was present at the fimbriated end of left fallopian tube involving 1 plica. It comprised crowded glandular proliferation with a pseudostratified columnar lining. The cells displayed elongated nuclei with no remarkable nuclear atypia.Immunohistochemistry showed patchy loss of PAX 2 expression with multifocal aberrant nuclear and cytoplasmic staining for B-catenin. p53 was wild-type and ER was positive.In view of the co-existing endometrioid type endometrial carcinoma, a possible metastatic spread to the fallopian tube was considered. However, morphologically no obvious nuclear atypia noted, and no associated inflammatory response or desmoplastic stromal reaction identified within the tubal lesion. And on immunostaining, the endometrial tumour was distinct from the tubal lesion. For instance, PTEN was negative/lost in the endometrial tumour but retained in the tubal lesion and B-catenin was membranous in the endometrial tumour but aberrant with multifocal nuclear and cytoplasmic overexpression in the tubal lesion. WT1 was negative in the endometrial tumour but positively expressed by the tubal lesion. All the above findings favoured the possibility of the tubal lesion as being independent of the endometrial primary. In conclusion, we describe an incidental B-catenin aberrant endometrioid type proliferation of the fallopian tube/E-TIN, to raise awareness of such lesions.

Activation of the WNT4/ β-catenin/FOXO1 pathway by PDK1 promotes cervical cancer metastasis and EMT process

This study aimed to elucidate the role of pyruvate dehydrogenase kinase-1 (PDK1) in cervical cancer (CC) by investigating its impact on cell proliferation, migration, and epithelial-mesenchymal transition (EMT) under hypoxic conditions. PDK1-silenced CC cell lines were established using lentiviral shRNA technology. Cell migration and invasion were assessed through scratch and Transwell assays, respectively. Cellular activity and apoptosis-related protein expression levels were evaluated using MTT assays and western blotting. Transcriptome sequencing elucidates the regulatory pathways impacted by PDK1 silencing, and rescue experiments confirmed the underlying mechanisms. Xenograft models with nude mice were used to validate the effects of PDK1 silencing on CC progression. PDK1 silencing reduced migration, invasion, and cellular activity under hypoxic conditions while promoting apoptosis. Transcriptomic analysis revealed that PDK1 suppression downregulated the WNT4/β-catenin/FOXO1 pathway, decreasing EMT-related protein expression. Mechanistically, PDK1 enhanced β-catenin stability by inhibiting its phosphorylation through AKT-mediated GSK3β inactivation, promoting EMT and anti-apoptotic gene transcription. Targeting PDK1 may provide novel therapeutic strategies specifically for CC by modulating the WNT4/β-catenin/FOXO1 pathway and associated EMT and apoptotic processes.

Upregulation of NR4A3 increases sensitivity to niraparib in ovarian cancer cells resistant to this drug

Acquired resistance is the major clinical change for patients with ovarian cancer undergoing niraparib (NRP) treatment. Targeting NRP-resistant cells would be NRP offers an effective strategy for reversing resistance and inhibiting disease progression. NR4A3 is known as a tumor suppressor in multiple cancers, but its role in NRP resistance of ovarian cancer is still unclear. NRP-resistant ovarian cancer cell lines (A2780/NRP and OVCAR3/NRP) were established by gradually increasing NRP concentrations. The phenotype of resistant cell lines was characterized using proliferation, colony formation, and migration assays. RNA sequencing was performed to identify genes dysregulated in the resistant cell lines. We also performed gain- and loss-of-function assays to investigate the role of NR4A3 in NRP resistance. The resistance indices for A2780/NRP and OVCAR3/NRP were 8.95 and 4.42, respectively. These resistant cells exhibited slower proliferation and robust colony formation and migration abilities. NR4A3 exhibited the highest average log2 fold change among the candidates. Overexpression of NR4A3 sensitized the NRP-resistant cells and reduced their proliferation, colony formation, and migration capabilities, whereas downregulation of NR4A3 in the parental cells caused opposite results. Downregulation of NR4A3 contributes to NRP resistance, and activation of NR4A3 maybe a promising strategy to reverse NRP resistance.

BRAF mutations in ovarian cancer: immune microenvironment and therapeutic advances

Ovarian cancer is a highly lethal malignancy of the female reproductive system, characterized by a poor prognosis and complex therapeutic challenges influenced by various molecular and microenvironmental factors. Among these, mutations in the BRAF gene have received increasing attention across multiple malignancies due to their involvement in oncogenic signaling pathways and implications for targeted therapy. Concurrently, the tumor immune microenvironment has been recognized as a critical determinant in the initiation and progression of tumors. This review explores the impact of BRAF mutations on the immune microenvironment in ovarian cancer, summarizes recent advances in the field, and assesses the potential clinical significance of these findings in informing therapeutic strategies. Clinical trial number: Not applicable.

Effect of sufentanil on the proliferation, apoptosis, and epithelial-mesenchymal transition of ovarian cancer cells by regulating the SMAD3/SNAIL signaling pathway

To study sufentanil's effect on the proliferation, apoptosis, and epithelial-mesenchymal transition (EMT) of ovarian cancer cells by modulating the SMAD3/SNAIL signaling pathway. Ovarian cancer A2780 cells were exposed to sufentanil at varying concentrations of 0 ng/mL, 20 ng/mL, 40 ng/mL, 60 ng/mL, 80 ng/mL, and 100 ng/mL. The proliferation of A2780 cells was assessed using the CCK-8 method; A2780 cells were randomly divided into three groups: CON group (normal culture), SUF group (60 ng/mL sufentanil intervention), and SUF + ACA group (60 ng/mL sufentanil and 100 ng/mL activin A intervention). After 48 h of culture, cell migration was evaluated by the scratch assay; Cell invasion was assessed using the Transwell chamber assay; Cell apoptosis was measured via flow cytometry; The growth status of the cells was observed under an optical microscope; The expression of N-cadherin, E-cadherin, SMAD3, TGF-β, and SNAIL proteins was detected by Western blot. When the concentration of sufentanil was ≥ 20 ng/mL, it dose-dependently inhibited the proliferation of ovarian cancer A2780 cells; In comparison to the CON group, the number of A2780 cells in the SUF group was significantly reduced, some cells detached, cell migration and invasion abilities, and the expression levels of N-cadherin, SMAD3, TGF-β, and SNAIL proteins decreased, while the apoptosis rate and E-cadherin protein expression levels increased (P < 0.05); In comparison to the SUF group, the growth status of A2780 cells in the SUF + ACA group was good, cell migration and invasion abilities, and the expression levels of N-cadherin, SMAD3, TGF-β, and SNAIL proteins increased, while the apoptosis rate and E-cadherin protein expression levels decreased (P < 0.05). Sufentanil may inhibit the proliferation and epithelial-mesenchymal transition of ovarian cancer cells and promote cell apoptosis by inhibiting the SMAD3/SNAIL signaling pathway.

4SC-202 exerts an anti-tumor effect in cervical cancer by targeting PRLR signaling pathway

The aim of the present study is to investigate whether 4SC-202, a selective class I histone deacetylase inhibitor (HDACi), plays an anti-tumor role in cervical cancer (CC) by targeting prolactin receptor (PRLR). CCK-8 and colony formation assays were used to evaluate the effects of 4SC-202 on the proliferation of CC cells in vitro. Effects of 4SC-202 on the cell cycle distribution and apoptosis in SiHa cells were determined by flow cytometry and western blotting, respectively. Immunofluorescence, western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to detect the activities of PRLR-related pathways and PRLR expression in CC cells. A xenograft tumor model in nude mice was established to examine effects of 4SC-202 on the tumor growth, apoptosis and PRLR-related pathways in vivo. The biochemical analyzer and H&E staining were used to detect the serum biochemical indexes and organ toxicity. 4SC-202 inhibited the proliferation of CC cells (SiHa, HeLa, and CaSki) in vitro in a time- and dose-dependent manner. SiHa cells were treated with 1 or 5 µM 4SC-202 for 72 h and then subjected to various functional assays. The assays showed that 4SC-202 significantly induced G2/M phase arrest and apoptosis, while inhibiting the activities of PRLR-related pathways and PRLR expression. In addition, 4SC-202 reduced tumor growth and induced apoptosis in vivo. 4SC-202 down-regulated the expression of PRLR and activities of PRLR-related pathways in the mouse model, displayed no effects on serum biochemical indicators and caused no toxicity to mouse organs. This finding suggests that 4SC-202 may serve as a novel therapeutic agent for CC.

Adipose-derived stem cell exosomes promote endometrial carcinoma progression via MAGED4B/CDH1/EMT axis

Adipose-derived stem cell exosomes (ADSCs-Exos) enhance endometrial carcinoma (EC) cell proliferation, migration, and invasion. Mechanistically, ADSCs-Exos downregulate MAGED4B and CDH1, reduce E-cadherin expression, and upregulate vimentin, promoting epithelial-mesenchymal transition (EMT). Overexpression of MAGED4B reverses these effects and inhibits malignant behaviors. Furthermore, ADSCs-Exos increase organoid viability and confirm key protein changes. These findings demonstrate that ADSCs-Exos promote EC progression via the MAGED4B/CDH1/EMT axis.

SR-B1 and CD10 combined immunoprofile for differential diagnosis of metastatic clear cell renal cell carcinoma and clear cell carcinoma of the ovary

AbstractBoth clear cell renal carcinoma (ccRCC) and clear cell carcinoma of the ovary (CCOC) have a clear cytoplasmic morphological feature, hence it is difficult to identify metastatic ccRCC and CCOC by morphology alone. At present, there are no effective immunohistochemical markers to distinguish between these two tumors. Studies have shown that the clear cytoplasm of ccRCC is mainly caused by cholesterol-rich lipids in the cytoplasm, while that of CCOC is due to the accumulation of cytoplasmic glycogen. Objective: to hypothesize that the scavenger receptor class B-type 1 (SR-B1) protein responsible for HDL cholesterol uptake may be differentially expressed in ccRCC and CCOC, and high CD10 expression in the renal tubular epithelium may assist in distinguishing between ccRCC and CCOC. Methods: effective immunohistochemical markers were applied in 90 cases of renal clear cell carcinoma and 31 cases of ovarian cancer to distinguish between the two types of tumors.Result: SR-B1 and CD10 expression is significantly higher in ccRCC than CCOC. Both SR-B1 and CD10 exhibited focal weak-medium intensity staining in CCOC, and their staining extent and intensity were significantly lower than ccRCC. The sensitivity and specificity of SR-B1 for identifying ccRCC were 74.4% and 83.9%, respectively. The sensitivity and specificity of CD10 for identifying CCOC were 93.3% and 80.6%, respectively. The combined SR-B1( +) CD10( +) immunoprofile supports the diagnosis of ccRCC with a specificity of 93.5%. The combined SR-B1(-) CD10(-) immunoprofile supports the diagnosis of CCOC with a specificity of 93.3%. Conclusions: our findings demonstrate that the combination of SR-B1 and CD10 immunoprofiling is a valuable tool for differential diagnosis of ccRCC and CCOC.

Long noncoding RNA HOXC-AS3 enhances the progression of cervical cancer via activating ErbB signaling pathway

Emerging evidence reveals that long noncoding RNAs (lncRNAs) contribute to human tumorigenesis. Nevertheless, the function of HOXC cluster antisense RNA 3 (HOXC-AS3) in human cervical cancer (CC) remains largely unknown. The levels of HOXC-AS3, miR-105-5p and SOS1 in CC tissues and cells were monitored by reverse transcription-polymerase chain reaction (RT-PCR) and western blot (WB). Gain- and loss-of-function experiments were conducted to verify the function of HOXC-AS3 and miR-105-5p in CC cells. Meanwhile, cell proliferation, apoptosis, migration and invasion were examined by the cell counting kit-8 (CCK8) experiment, colony formation assay, flow cytometry and Transwell assay. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were carried out to test the regulatory interaction of HOXC-AS3, miR-105-5p and SOS1. In addition, in vivo experiment was performed to certain the role of HOXC-AS3 in tumorigenesis of CC. HOXC-AS3 was overexpressed in CC tissues (vs. adjacent normal tissues) and CC cells. Besides, the higher HOXC-AS3 profile was associated with the poorer clinical prognosis of CC patients. Overexpression of HOXC-AS3 promoted cell growth, migration and invasion, hampered apoptosis, whereas knocking down HOXC-AS3 exhibited the reverse effects. MiR-105-5p was a downstream target of HOXC-AS3, and it mediated the HOXC-AS3-induced oncogenic effects. Mechanistically, the bioinformatic analysis illustrated that SOS1 was targeted by miR-105-5p. Up-regulating SOS1 heightened the growth, migration and invasion of CC cells by enhancing the ErbB signaling pathway, which was reversed by miR-105-5p. Up-regulated HOXC-AS3 aggravates CC by promoting SOS1 expression via targeting miR-105-5p.

MiR-135b improves proliferation and regulates chemotherapy resistance in ovarian cancer

MicroRNAs act as regulators in ovarian tumorigenesis and progression by involving different molecular pathways. Here, we examined the role of miR-135b on growth, chemotherapy resistance in OVCAR3 and SKOV3 ovarian cancer cells. MTT assay was performed to examine proliferation. Transwell migration and matrigel invasion assays were used to assess migration and invasion. Caspase-Glo3/7 assay was carried out to evaluate apoptosis. The dual-luciferase reporter assay was performed to validate the putative binding site. Meanwhile, the miR-135b levels in human ovarian cancer tissue were detected by qPCR assay. Overexpression of miR-135b increased growth, and improved migration and invasion in ovarian cancer cells. Meanwhile, overexpression of miR-135b decreased the cisplatin treatment sensitivity in OVCAR3 and SKOV3 cells. The cisplatin-induced apoptosis was decreased by miR-135b. Furthermore, miR-135b could alter epithelial to mesenchymal transition (EMT) associated proteins expression including E-cadherin, N-cadherin, snail and Vimentin in ovarian cancer cells. Further study demonstrated aberrant expression of miR-135b regulated PTEN and p-AKT expression in ovarian cancer cells. The expression level of miR-135b was increased in human ovarian cancer tissue, compared with normal ovary tissue. MiR-135b involves in tumorigenesis and progression in ovarian cancer cells, and might serve as a promising biomarker to predict chemotherapy sensitivity and prognosis in ovarian cancer.

Echinacoside inhibits the proliferation, migration, invasion and angiogenesis of ovarian cancer cells through PI3K/AKT pathway

Echinacoside is a group of natural compounds extracted from medicinal plants Cistanche and Echinacea, which has neuroprotective, antiaging, immunomodulatory and anticancer effects, but its specific role and mechanism in tumor remains partially unclear. To our knowledge, it was the first time to reported the effect of Echinacoside in ovarian cancer. Colony formation, TUNEL staining, Transwell and tube formation assays were conducted to analyze the proliferation, apoptosis, invasion and tube formation abilities of serous ovarian carcinoma cells (SKOV3 and OVCAR-3), respectively. The expressions of apoptosis-, invasion- and PI3K/AKT pathway-related proteins were measured by western blotting. In addition, PI3K agonist (740Y-P) was used to assess the regulatory effect of Echinacoside on PI3K/AKT signaling in ovarian cancer. Finally, the anti-tumor effect of Echinacoside on SKOV3-xenografted mice was evaluated by xenograft tumor mouse model. Our results demonstrated Echinacoside concentration-dependently reduced the proliferation, migration and angiogenesis of ovarian cancer cells, whereas promoted apoptosis. Moreover, western blotting revealed that Echinacoside suppressed the growth of ovarian cancer cells by downregulating the phosphorylation levels of PI3K, AKT and mTOR, which could be partially reversed by 740Y-P. Further, in vivo results showed that Echinacoside could effectively alleviate the tumor growth of xenograft mice, accompanied by the decrease of PI3K/AKT signaling. In general, our results demonstrate that Echinacoside could reduce the ovarian cancer progression through inhibition of PI3K/AKT pathway, suggesting that Echinacoside may be a new treatment option for ovarian cancer.

Expression of cancer susceptibility candidate 11 in ovarian cancer tissues and its role in doxorubicin resistance

We aimed to investigate the expression of cancer susceptibility candidate 11 (CASC11) in ovarian cancer (OC) tissues and its role in doxorubicin (Dox) resistance. A total of 98 patients were included as subjects. Reverse transcription-polymerase chain reaction was employed to determine the expressions of CASC11 in OC and para-OC tissues, and in OC cells (A2780, SKOV3, OVCAR3 and A547) and human normal ovarian epithelial cells (IOSE-80) from these patients. OC SKOV3/R cell line with Dox resistance was established and transfected with small interfering (si)-CASC11 to down-regulate CASC11 expression. Based on the constructed nude mouse model of orthotopic transplanted tumor, the growth curves were plotted, and the changes in tumor volume and apoptosis were observed by hematoxylin-eosin staining. OC tissues had a significantly higher mRNA expression of CASC11 than that of para-OC tissues (P < 0.05). A547, OVCAR3, A2780 and SKOV3 cells had significantly higher mRNA expressions of CASC11 than that of IOSE-80 cells (P < 0.05). The transplanted tumor was significantly smaller in volume in the si-CASC11 group than that in the si-normal control (NC) group from the 8th days after transplanted tumor inoculation (P < 0.05). The tumor growth inhibition rate significantly rose in the si-CASC11 group in comparison with that in the si-NC group (P < 0.05). CASC11 has high expression in OC tissues. Knockout of CASC11 weakens the proliferative, invasive and migratory potentials and enhances the apoptotic potential of Dox-resistant OC cells, thereby reversing their Dox resistance.

Morin induces ferroptosis in endometrial cancer cells by down-regulating FTH1

Ferritin heavy chain 1 (FTH1) is abnormally expressed in various cancers, but its role and mechanism in endometrial cancer (EC) remain unclear. This study aims to explore the clinical significance, biological functions and potential inhibitor of FTH1 in EC. The expression, prognosis and clinical correlation of FTH1 in EC were analyzed using TIMER, GEO, kaplan-meier plotter and UALCAN databases. Virtual screening and molecular docking were conducted for identifying potential inhibitors of FTH1. In vitro experiments were conducted using human EC cells HEC-1A and RL95-2. Cell proliferation, cell cycle and apoptosis were detected by CCK-8 assay and flow cytometry. The level of reactive oxygen species (ROS) was detected by using the DCFH-DA probe. The levels of malondialdehyde (MDA) and glutathione (GSH) were detected using the corresponding test kits. Western blot was used to detect the expression level of FTH1, AKT and p-AKT. FTH1 was highly expressed in EC tissues and was associated with a shorter overall survival time of patients. Functional enrichment analysis revealed that FTH1 was mainly involved in the iron homeostasis and ferroptosis pathways. FTH1 knockdown inhibits the proliferation of EC cells, induces cell cycle arrest at the G0/G1 phase and triggers cell apoptosis. In EC cells with FTH1 knockdown, the levels of ROS and MDA were significantly increased, accompanied by a decrease in GSH levels. Furthermore, morin had a high binding affinity with FTH1, which also inhibited the malignant phenotypes of EC cells. Morin triggered ferroptosis of EC cells by down-regulating FTH1 expression and inhibiting the PI3K/AKT pathway. FTH1 is a potential prognostic biomarker and therapeutic target for EC. Morin induces ferroptosis of EC cells by regulating FTH1-PI3K/AKT axis, providing a new candidate drug and theoretical basis for the treatment of EC.

LINC02418 suppresses endometrial cancer progression via regulating miR-494-3p/RASGRF1 axis

Long non-coding RNAs (lncRNAs) have emerged as pivotal regulatory molecules in cancer biology. Among these, long intergenic non-protein coding RNA 02418 (LINC02418), a recently identified lncRNA, has been linked to endometrial cancer (EC), although its function and operational mechanisms are largely unclear. The present investigation aims to elucidate the molecular mechanism through which LINC02418 influences EC pathogenesis. We employed Western blotting and quantitative real-time PCR to analyze Ras protein specific guanine nucleotide releasing factor 1 (RASGRF1) and LINC02418 expression profiles in EC tissues and cell lines. Functional analyses, including cell proliferation, migration, and invasion assays, were conducted to evaluate the impact of LINC02418 overexpression on EC cells. Xenograft mouse models were established for in vivo validation. The molecular interactions between LINC02418, miR-494-3p, and RASGRF1 were characterized using luciferase reporter and RNA pull-down assays. LINC02418 expression was significantly downregulated in EC tissues and cell lines compared to their normal counterparts. Forced expression of LINC02418 significantly suppressed EC cell proliferation, migration, and invasion in vitro. In xenograft models, LINC02418 overexpression resulted in reduced tumor burden and enhanced cell death. Mechanistically, LINC02418 enhanced RASGRF1 expression by sequestering miR-494-3p, a finding substantiated by RNA pull-down assays. The tumor-suppressive effects of LINC02418 were partially reversed by RASGRF1 silencing and miR-494-3p overexpression. Clinical analyses revealed that reduced RASGRF1 expression correlated with poor histological differentiation, advanced tumor stages, and decreased overall survival in EC patients. Our findings establish LINC02418 as a tumor suppressor that regulates EC progression through modulation of the miR-494-3p/RASGRF1 axis, highlighting its potential as a therapeutic target in EC treatment.

TFDP1 transcriptionally activates KIF22 to enhance aggressiveness and stemness in endometrial cancer: implications for prognosis and targeted therapy

This study aims to elucidate the role of Kinesin Family Member 22 (KIF22) as a critical regulator of aggressive behavior in endometrial cancer (uterine corpus endometrial carcinoma, UCEC) and to uncover its underlying mechanisms, thereby providing a molecular rationale for future targeted treatment. Bioinformatics analyses were employed to assess KIF22 and TFDP1 expression in UCEC, examining their prognostic value and associations with disease progression. Expression levels were validated in UCEC tissues using qRT-PCR and western blotting. Potential TFDP1 binding sites on the KIF22 promoter were predicted using the JASPAR database and confirmed via dual-luciferase reporter assays. Functional assays, including CCK-8, transwell, and spheroid formation assays, were conducted to evaluate the effects of KIF22 knockdown on UCEC cell behavior. A mouse xenograft model was utilized to investigate the in vivo impact of KIF22 suppression on tumor growth and stemness. KIF22 expression was significantly elevated in UCEC tissues, correlating with reduced overall survival in patients with high KIF22 levels. Overexpression of KIF22 enhanced the proliferation, migration, and sphere formation of UCEC cells. Similarly, high TFDP1 expression was associated with poorer patient outcomes. KIF22 was found to be positively regulated by the TFDP1 transcription factor, which bound to the KIF22 promoter and activated its expression in UCEC cells. In vivo, KIF22 knockdown markedly impeded the tumor formation of cells and reduced stemness marker expression. KIF22, upregulated by TFDP1, enhances UCEC cell aggressiveness and is linked to poor prognosis, highlighting its potential as a target for therapeutic intervention in endometrial cancer.