HO-8910 Cells Research Articles

The possible regulatory effect of the PD-1/PD-L1 signaling pathway on Tregs in ovarian cancer.

Aim of this study was to investigate the possible regulatory effect of the programmed death-1 (PD-1)/programmed death ligand-1 (PD-L1) signaling pathway on Tregs in ovarian cancer. Immunohistochemistry was used to detect the expression of PD-L1 and PD-1 and the presence of FOXP3+ Tregs in ovarian cancer. Then, ovarian cancer HO8910 cells were subjected to transfection with PD-L1 siRNA in vitro. CCK-8, Transwell and wound healing assays were performed to detect the biological behaviors of ovarian cancer cells. Human T-cells isolated from human peripheral blood were cocultured with HO8910 cells, which were divided into the Control, TGF-β, and TGF-β+ anti-PD-L1 groups. The proportion of differentiated Tregs was detected by flow cytometry. Mouse models of ovarian cancer were established, and PD-L1 antibody therapy was administered. Tumor growth and Treg recruitment were observed. PD-L1, PD-1 and FOXP3+ Tregs were found in ovarian cancer tissue. Patients with tumors with an advanced stage and low differentiation and lymph node metastasis had significantly higher levels of PD-1, PD-L1 and FOXP3+ Tregs. After transfection with PD-L1 siRNA, HO8910 cells showed a significant reduction in PD-L1 expression, proliferation, migration and invasion. After T-cells were cocultured with ovarian cancer cells, the TGF-β+ anti-PD-L1 group showed a substantial decline in the differentiation of T-cells into Tregs compared with the TGF-β group. Moreover, mice in the anti-PD-L1 group had significantly reduced tumor growth rates, Treg proportions in the tumor microenvironment, and FOXP3 expression.

Overexpression of DUSP6 enhances chemotherapy-resistance of ovarian epithelial cancer by regulating the ERK signaling pathway.

Objective: DUSP6 is a negative regulator of the ERK signaling pathway and plays an important role in chemotherapy-resistance. Previously we showed that DUSP6 is overexpressed in ovarian cancer side population (SP) cells that possess cancer stem cell-like properties and are quiescent and chemotherapy-resistant. Here, we explore the effects of DUSP6 on chemotherapy-resistance by examining its regulation of the ERK signaling pathway and G0/G1 cell cycle arrest.Methods: mRNA and protein expression of DUSP6 and G0/G1 cell cycle checkpoint regulating proteins (CyclinD1, CyclinD3 and CyclinE2) was evaluated among ovarian cancer cell lines and tissue samples. Ovarian cancer cells were transiently transfected to overexpress DUSP6. After treatment with cisplatin, cell viability was measured by the MTS assay at 48 hours and the half maximal inhibitory concentration (IC50) for each cell line was calculated. Subcellular localization and cell cycle analysis were determined by using immunofluorescence and FACS, respectively.Results: SKOV3 and OVCAR8 SP cells were shown to express higher levels of DUSP6 and lower levels of CyclinD3 compared with non-SP (NSP) cells (P<0.001). Among 39 ovarian cancer tissue samples, expression of DUSP6 in the chemotherapy-resistant group (12 samples) was higher than in the chemotherapy-sensitive group (27 samples) (P<0.05). While a lower level of expression of CyclinD3 was seen in the chemotherapy-resistant group, it was not statistically different from the chemotherapy-sensitive group. HO8910 cells where shown to have higher IC50 to cisplatin than SKOV3 or OVCAR8 cells, and this correlated with higher levels of DUSP6 expression. Overexpression of DUSP6 in SKOV3 cells led to an increase in cisplatin IC50 values (P<0.05), and also markedly reduced the expression levels of phospho-ERK1/2 and CyclinD3 and to the predominance of cells in the G0/G1 phase.Conclusion: Our findings reveal an enhancement of chemotherapy-resistance and a predominance of cells in G1 cell cycle arrest in DUSP6-overexpressing ovarian cancer cells. This suggests that overexpression of DUSP6 promotes chemotherapy-resistance through the negative regulation of the ERK signaling pathway, increasing the G0/G1 phase ratio among ovarian cancer cells, and leading to cellular quiescence.

CHD1L promotes EOC cell invasiveness and metastasis via the regulation of METAP2.

Chromodomain helicase DNA binding protein 1-like (CHD1L) gene has been proposed to play an oncogenic role in human hepatocellular carcinoma. Previously we reported that CHD1L overexpression is significantly associated with the metastasis proceeding of epithelial ovarian cancer (EOC), and may predict a poor prognosis in EOC patients. However, the potential oncogenic mechanisms by which CHD1L acts in EOC remain unclear. To elucidate the oncogenic function of CHD1L, we carried out a series of in vitro assays, with effects of CHD1L ectogenic overexpression and silencing being determined in EOC cell lines (HO8910, A2780 and ES2). Real-time PCR and Western blotting analyses were used to identify potential downstream targets of CHD1L in the process of EOC invasion and metastasis. In ovarian carcinoma HO8910 cell lines, ectopic overexpression of CHD1L substantially induced the invasive and metastasis ability of the cancer cells in vitro. In contrast, knockdown of CHD1L using shRNA inhibited cell invasion in vitro in ovarian carcinoma A2780 and ES2 cell lines. We also demonstrated that methionyl aminopeptidase 2 (METAP2) was a downstream target of CHD1L in EOC, and we found a significant, positive correlation between the expression of CHD1L and METAP2 in EOC tissues (P<0.05). Our findings indicate that CHD1L plays a potential role in the inducement of EOC cancer cell invasion and/or metastasis via the regulation of METAP2 expression and suggests that CHD1L inhibition may provide a potential target for therapeutic intervention in human EOC.

LncRNA DANCR aggravates the progression of ovarian cancer by downregulating UPF1.

To uncover the role of long non-coding RNA (lncRNA) DANCR in aggravating the progression of ovarian cancer (OC) by downregulating UPF1 level. DANCR level in OC tissues and matched adjacent normal ones was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Its expression level in OC patients with different tumor node metastasis (TNM) staging and either with metastasis, or not, was examined as well. Receiver operating characteristic (ROC) curves were introduced for assessing the prognostic value of DANCR in OC. Subsequently, regulatory effects of DANCR on proliferative and migratory abilities of HO8910 and HEY cells were evaluated. Subcellular distribution of DANCR in OC cells was analyzed. Furthermore, the interaction between DANCR and UPF1 was explored by RNA immunoprecipitation (RIP) and Pearson correlation analysis. Finally, rescue experiments were conducted to clarify the role of DANCR/UPF1 axis in the progression of OC. DANCR was upregulated in OC tissues and cell lines. Its level was higher in OC patients with worse tumor stage and accompanied by metastatic loci. DANCR exerted the potential to serve as a prognostic marker for OC. Overexpression of DANCR accelerated HO8910 and HEY cells to proliferate and migrate. UPF1 was found to be downregulated in OC tissues and negatively correlated to DANCR. DANCR was mainly distributed in the cytoplasm and interacted with UPF1. Overexpression of UPF1 in OC cells partially reversed the promotive effect of DANCR on proliferative and migratory rates. LncRNA DANCR accelerates the proliferative and migratory abilities of OC cells through negatively regulating UPF1 level, thus aggravating the progression of OC.

MiR-203 regulates proliferation and apoptosis of ovarian cancer cells by targeting SOCS3.

Cytokine signal transduction inhibitor 3 (SOCS3) negatively regulates Janus kinases (JAK) - signal transducer and activator of transcription (STAT) pathway. Bioinformatics analysis revealed a targeted relationship between miR-203 and SOCS3 mRNA. This study investigated the role of miR-203 in ovarian cancer cell proliferation and apoptosis. Ovarian cancer tissues and adjacent tissues were collected to detect the expression of miR-203 and SOCS3. Ovarian cancer HO8910 cells were divided into miR-NC group, miR-203 inhibitor group, and miR-203 mimic group followed by the analysis of the expression of miR-203 and SOCS3 mRNA by quantitative Reverse Transcription-PCR (qRT-PCR), protein expression of p-JAK2 and p-STAT3 by Western blot, cell apoptosis by flow cytometry, and proliferation by 5-Ethynyl-2'-deoxyuridine (EdU) staining chronologically. Compared with adjacent tissues, miR-203 expression was significantly increased in tumor tissues and SCOS3 mRNA expression was decreased. Compared with those with lower miR-203 expression, the prognosis of patients with higher expression of miR-203 was significantly worse. There was a targeted regulatory relationship between miR-203 and SOCS3 mRNA. Compared with IOSE80 cells, miR-203 expression in HO8910 and SKOV3 cells was increased, and its expressions of SOCS3 mRNA and protein were decreased. Compared with miR-NC group, the transfection of miR-203 inhibitor significantly increased SOCS3 expression, and decreased the expression of p-JAK2 and p-STAT3 protein. We draw the conclusion that miR-203 increased cell apoptosis and decreased cell proliferation. However, opposite results were observed after the transfection of miR-203 mimic. Abnormal miR-203 and SOCS3 expression are related to the pathogenesis of ovarian cancer. MiR-203 affects the proliferation of JAK-STAT pathway and regulates the proliferation and apoptosis of ovarian cancer cells by targeting the inhibition of SOCS3 expression.

Upregulation of circular RNA circ‑FAM53B predicts adverse prognosis and accelerates the progression of ovarian cancer via the miR‑646/VAMP2 and miR‑647/MDM2 signaling pathways.

Ovarian cancer (OC) is a common cancer of the human genital system. Circular RNAs (circRNAs) play an important role in carcinogenesis and progression of various cancers. The present study aimed to clarify the expression profile and functions of circ‑FAM53B in the progression of OC and reveal its underlying mechanisms. Relative levels of circ‑FAM53B in OC specimens and cell lines were determined by quantitative real‑time polymerase chain reaction (qRT‑PCR). The clinical significance of circ‑FAM53B in OC patients was analyzed through Fisher's exact test, Kaplan‑Meier curves, and Cox regression analysis. Subsequently, the regulatory effects of circ‑FAM53B on the proliferative, apoptotic, migratory, and invasive potential of OC cells were determined by loss/gain‑of‑function experiments. Mechanistically, bioinformatics analysis and luciferase reporter gene assay were used to reveal the potential molecular mechanisms of circ‑FAM53B in OC. circ‑FAM53B was overexpressed in OC specimens and cells and correlated with clinical severity and poor prognosis of OC patients. The overexpression of circ‑FAM53B accelerated the proliferation, migration, and invasion of HO8910 cells; however, it decreased the number of apoptotic cells. Silencing of circ‑FAM53B induced the opposite effect. Through dual‑luciferase reporter gene assay and functional experiments, the potential functions of circ‑FAM53B/miRNA‑646/vesicle‑associated membrane protein 2 (VAMP2) and circ‑FAM53B/miRNA‑647/mouse double minute 2 (MDM2) in mediating the progression of OC were identified. Collectively, the present results indicated that circ‑FAM53B could be a competing endogenous RNA (ceRNA) to competitively sponge miR‑646 and miR‑647 to upregulate VAMP2 and MDM2 expression at the post‑transcriptional level, thus mediating the cellular behaviors of OC cells.

LINC01342 promotes the progression of ovarian cancer by absorbing microRNA-30c-2-3p to upregulate HIF3A.

Ovarian cancer (OC) is a highly prevalent gynecologic malignancy and its mortality is extremely high. Therefore, the development of novel therapeutic approaches for OC is of great significance. In this study, LINC01342 was upregulated in OC tissue in the GSE38666 microarray and in tumor tissue samples collected in our center. The silencing of LINC01342 suppressed the proliferative and metastatic capacities of A2780 and HO8910 cells. Subcellular distribution assays showed that LINC01342 was mainly enriched in the cytoplasm. Subsequently, the downregulation of microRNA-30c-2-3p was proven to be the target of LINC01342. The silencing of microRNA-30c-2-3p enhanced the clonality and migratory capacity of OC cells. Moreover, the silencing of microRNA-30c-2-3p could reverse the inhibited migration and clonality in OC cells caused by LINC01342 knockdown. In addition, hypoxia-inducible factor 3 subunit α (HIF3A) was proven to be the target gene of microRNA-30c-2-3p, which was upregulated. HIF3A was negatively regulated by microRNA-30c-2-3p but positively regulated by LINC01342 in OC cells. An RNA binding protein immunoprecipitation assay showed that microRNA-30c-2-3p, LINC01342, and HIF3A could bind to argonaute RISC catalytic component 2. The overexpression of HIF3A reversed the inhibited migration and clonality in OC cells with LINC01342 knockdown. By analyzing the follow-up data from the enrolled OC patients, the LINC01342 and HIF3A levels were negatively correlated with prognosis, while the microRNA-30c-2-3p level was positively correlated with the same. In short, the upregulated LINC01342 in OC absorbs microRNA-30c-2-3p to release HIF3A. Thus, upregulated HIF3A expression accelerates the progression of OC.

LINC01210 accelerates proliferation, invasion and migration in ovarian cancer through epigenetically downregulating KLF4

BackgroundIn recent years, a large number of studies have shown that differentially expressed lncRNAs in tumors are capable of inducing tumorigenesis and promoting tumor development. However, the role of LINC01210 in OC still remains unclear. MethodsRelative levels of LINC01210 and KLF4 in OC tissues containing in the GSE40595 dataset and those collected in our hospital were determined. Prognostic potential of LINC01210 in OC was evaluated by Kaplan-Meier method. Correlation between expression levels of LINC01210 and KLF4 was assessed by Spearman correlation test. After silence of LINC01210 in A2780 and HO8910 cells, changes in proliferative, migratory and invasive abilities were examined. Moreover, the interaction between LINC01210 and KLF4 was explored by RIP and ChIP. Rescue experiments were conducted to uncover the involvement of KLF4 in LINC01210-regulated OC progression. ResultsLINC01210 was upregulated in OC tissues and KLF4 was downregulated. LINC01210 level was higher in OC patients with metastases or advanced stage. Besides, its level was negatively correlated to DFS (disease-free survival) and OS (overall survival) of OC patients. Silence of LINC01210 attenuated proliferative, migratory and invasive abilities in A2780 and HO8910 cells. Through analyzing the GSE40595 dataset, LINC01210 was found to be negatively linked to KLF4. RIP assay further verified the interaction between LINC01210 and KLF4. Knockdown of LINC01210 markedly decreased the recruitment ability of EZH2 to KLF4. Importantly, silence of KLF4 could reverse regulatory effects of LINC01210 on cellular behaviors of OC. ConclusionsLINC01210 is upregulated in OC and predicts a poor prognosis. It accelerates proliferation, invasion and migration in OC cells through epigenetically downregulating KLF4.

Long non-coding RNA LINC01627 is a prognostic risk factor for epithelial ovarian cancer.

Ovarian malignancies are commonly diagnosed cancers of the female reproductive system. Recent studies have revealed that long non-coding RNAs (lncRNAs) can regulate a variety of oncological processes. In the present study, ovarian cancer expression datasets were searched in the GEO database using the GPL570 platform. Differential lncRNA expression between normal ovarian tissues and ovarian tumors were analyzed using the R package ‘limma’, and patient prognosis was accessed using the package ‘survival’. Four databases, GSE14001, GSE18520, GSE38666 and GSE40595, were used for the analysis. A total of 64 lncRNAs were highly expressed and 4 were downregulated within these four databases. Prognostic analysis of the 68 lncRNAs in the four databases was performed, and revealed that the expression of long intergenic non-protein coding RNA 1627 (LINC01627) was negatively associated with patient prognosis in GSE19829 and GSE62193; there was no association between LINC01627 expression and patient's prognosis in GSE18520 or GSE63885. To investigate the proposed association between LINC01627 and patient prognosis, meta-analysis revealed that the total hazard ratio was 1.38 and the 95% confidence interval was between 1.04 and 1.83. Subgroup analysis revealed that LINC01627 may predict patient prognosis in high-grade, advanced and serous epithelial ovarian cancer, which was a risk factor for prognosis. Further assessment was performed in clinical samples and ovarian cancer cells, where the knockdown of LINC01627 inhibited the proliferative and migratory capacities of HO8910 and HEY cells. Collectively, the present results suggested that lncRNA LINC01627 may serve an oncogenic role in the development of epithelial ovarian tumors.

KCNMA1-AS1 attenuates apoptosis of epithelial ovarian cancer cells and serves as a risk factor for poor prognosis of epithelial ovarian cancer.

To explore the role of KCNMA1-AS1 in epithelial ovarian cancer (EOC) and its underlying mechanism. We first screened out the differentially expressed lncRNAs (KCNMA1-AS1) in the GEO (gene expression omnibus) database. The relationship between KCNMA1-AS1 expression and prognosis of EOC with different pathological types was analyzed by meta-analysis. Subsequently, KCNMA1-AS1 expressions in EOC tissues and normal ovarian tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between KCNMA1-AS1 level with progression-free survival (PFS) and overall survival (OS) of EOC was analyzed. Furthermore, proliferation and migration of EOC cells transfected with the corresponding plasmids were analyzed by Cell Counting Kit-8 (CCK-8) and transwell assay, respectively. Apoptosis-related genes in EOC cells were detected by Western blot. KCNMA1-AS1 was a risk factor for prognosis in high-grade, advanced and serous EOC. Upregulated KCNMA1-AS1 was found in EOC tissues than that of normal tissues, showing the diagnostic potential of KCNMA1-AS1 in EOC. Statistical analysis indicated that KCNMA1-AS1 was not correlated with the DFS, OS, age, histological type, lymph node metastasis and recurrence, but related to FIGO stage of EOC patients. For in vitro experiments, the proliferation and migration of were enhanced, and apoptosis of HO8910 cells overexpressing KCNMA1-AS1 was inhibited. Furthermore, elevated expressions of Caspase-3 and Caspase-9, as well as reduced expression of Bcl-xL, were observed after KCNMA1-AS1 knockdown in EOC cells. KCNMA1-AS1 is overexpressed in EOC and negatively correlated with its prognosis. KCNMA1-AS1 participates in the occurrence and development of EOC by promoting proliferation, migration and inhibiting apoptosis of ovarian cancer cells via apoptosis pathway.

LINC00968 accelerates the progression of epithelial ovarian cancer via mediating the cell cycle progression.

The aim of this study was to clarify the potential role of LINC00968 in the progression of epithelial ovarian cancer (EOC) and the underlying mechanism. The relative expression level of LINC00968 in EOC tissues (n=40) and normal ovarian tissues (n=40) was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). LINC00968 expression in human-derived ovarian cancer cell lines was examined by qRT-PCR as well. After transfection of LINC00968 small-interfering RNA (siRNA) in ovarian cancer cells, cell cycle progression and cell proliferation were evaluated through flow cytometry, Cell Counting Kit-8 (CCK-8) and colony formation assay, respectively. Tumor xenograft was conducted in nude mice to elucidate the function of LINC00968 in EOC tumorigenesis in vivo. Furthermore, the relative expression levels of cell cycle factors and protein kinase B/extracellular-signal-regulated kinase (AKT/ERK) in ovarian cancer cells influenced by LINC00968 were detected by Western blot. LINC00968 was significantly up-regulated in EOC tissues when compared with normal control tissues. Meanwhile, LINC00968 expression was positively correlated with the prognosis of EOC. Transfection of LINC00968 siRNA in HEY and HO8910 cells markedly attenuated proliferative ability and arrested cell cycle in the G1 phase. Knockdown of LINC00968 remarkably suppressed tumor growth of EOC in nude mice. The silence of LINC00968 significantly downregulated Cyclin D, Cyclin E and CDK4, whereas upregulated p16 and p21. In addition, AKT and ERK pathways were inhibited by knockdown of LINC00968 in ovarian cancer cells. LINC00968 expression is markedly upregulated in EOC. Meanwhile, it arrests the cell cycle in the G1 phase by inhibiting the ERK and AKT pathways, thus accelerating EOC progression.

New Cytotoxic Steroid Produced by the Soil-Derived Fungus Aspergillus flavus JDW-1

One new steroid 1, together with seven known compounds 2 to 8, were discovered in the extract of a soil-derived fungus Aspergillus flavus JDW-1. The structure, including the absolute stereochemistry of new compound 1, was established by interpretation of extensive nuclear magnetic resonance spectroscopic data and further confirmed by X-ray crystallographic analysis. The cytotoxicity of 1 against the A-549, Hela, HCT-116, MGC-803, and HO-8910 cell lines was evaluated, which showed cytotoxicity with the half-maximal inhibitory concentration (IC50) values as 5.00 to 22.38 μM. Compounds 2 to 8 exhibited moderate radical scavenging activity against DPPH with IC50 values ranging from 4.7 to 28.5 μM.

Discovery of HB-EGF binding peptides and their functional characterization in ovarian cancer cell lines

Ovarian cancer is one of the most frequent causes of cancer death among all gynecologic cancers. Though standard therapy often results in temporary clinical remission, most patients suffer from recurrence and metastasis of ovarian cancer, which highlights the need for developing new therapeutic agents targeting specific molecules. Previous studies have demonstrated that the native ligand of epidermal growth factor receptor (EGFR) and ErbB4, heparin-binding EGF-like growth factor (HB-EGF), plays a critical role in the progression of ovarian cancer and is associated with prognosis of ovarian cancer. In the current study, we tried to develop a peptide-based treatment for ovarian cancer by targeting HB-EGF. After the functions of HB-EGF in promoting migration and invasion of SKOV3 and HO-8910 cells were confirmed, phage display was used to discover peptides binding to HB-EGF. Two peptides, no. 7 and no. 29 were found mildly binding to HB-EGF. Then the effects of these peptides on HB-EGF functions were examined and both peptides no. 7 and no. 29 were found indeed inhibiting the functions of HB-EGF in promoting migration and invasion of SKOV3 and HO-8910 cells in vitro. Further mechanism investigation showed that peptides no. 7 and no. 29 inhibited HB-EGF-promoted cell migration and invasion through attenuating activation of the EGFR signaling pathway manifested by decreased p-Erk1/2 and Snail levels. More importantly, peptides no. 7 and no. 29 showed strong activities in inhibiting migration of SKOV3 cells in vivo. These results provide a proof of concept method for developing novel peptide drugs to combat ovarian cancer through interfering with HB-EGF mediated signaling pathways.

Ovarian Cancer Stem Cells with High ROR1 Expression Serve as a New Prophylactic Vaccine for Ovarian Cancer.

Tumor vaccines offer a number of advantages for cancer treatment. In the study, the vaccination with cancer stem cells (CSCs) with high expression of the type I receptor tyrosine kinase-like orphan receptor (ROR1) was evaluated in a murine model for the vaccine's immunogenicity and protective efficacy against epithelial ovarian carcinoma (EOC). CD117+CD44+ CSCs were isolated from human EOC HO8910 cell line using a magnetic-activated cell sorting system; murine ID8 EOC suspension sphere cells, which are collectively known as cancer stem-like cells, were acquired from serum-free suspension sphere-forming culture. Mice were subcutaneously immunized with the repeat cycles of freezing and thawing whole HO8910 CD117+CD44+ CSCs and ID8 cancer stem-like cells, respectively, followed by a challenge with HO8910 or ID8 cells at one week after final vaccination. The results showed that the CSC vaccination significantly induced immunity against EOC growth and markedly prolonged the survival of EOC-bearing mice in the prophylactic setting compared with non-CSC vaccination. Flow cytometry showed significantly increased immunocyte cytotoxicities and remarkably reduced CSC counts in the CSC-vaccinated mice. Moreover, the protective efficacy against EOC was decreased when the ROR1 expression was downregulated by shRNA in CSC vaccines. The findings from the study suggest that CSC vaccines with high ROR1 expression were highly effective in triggering immunity against EOC in vaccinated mice and may serve as an effective vaccine for EOC immunoprophylaxis.

Mechanism of Low Expression of miR-30a-5p on Epithelial-Mesenchymal Transition and Metastasis in Ovarian Cancer.

Metastasis of ovarian cancer is regulated by microRNAs. This study focused on the effects of miR-30a-5p on ovarian cancer migration and invasion. Our results showed that the miR-30a-5p and mucin type O-glycan biosynthesis are closely related to ovarian cancer, and that miR-30a-5p was downregulated in ovarian cancer cells. miR-30a-5p overexpression reduced cell viability and inhibited migration and invasion in HO-8910 and HO-8910PM cells. S phase kinase-associated protein 2 (SKP2), B cell lymphoma 9 (BCL9), and NOTHC1 are direct target genes of miR-30a-5p. MTDH, SKP2, BCL9, and NOTCH1 genes were overexpressed in ovarian cancer cells, and they are direct target genes of miR-30a-5p. miR-30a-5p overexpression inhibited epithelial-mesenchymal transition (EMT) process, while upregulation of SKP2, BCL9, and NOTCH1 gene expression levels reduced the inhibition of EMT process by miR-30a-5p. miR-30a-5p was lowly expressed in ovarian cancer, and such a phenomenon is related to ovarian cancer metastasis. miR-30a-5p might inhibit the migration and invasion of ovarian cancer cells by downregulating the expression of SKP2, BCL9, and NOTCH1 genes.

LncRNA SNHG12 accelerates the progression of ovarian cancer via absorbing miRNA-129 to upregulate SOX4.

To clarify whether long non-coding RNA (lncRNA) SNHG12 could regulate the proliferative and migratory abilities of ovarian cancer (OC) cells through mediating microRNA-129 (miRNA-129), thus influencing the progression of OC. The expression patterns of SNHG12 and miRNA-129 in OC tissues and adjacent normal tissues were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Meanwhile, their expression levels in OC cell lines were also examined. Regulatory effects of SNHG12 and miRNA-129 on the proliferative and migratory abilities of OC cells were evaluated by cell counting kit-8 (CCK-8) and transwell assay, respectively. Through the dual-luciferase reporter gene assay, we explored the binding between miRNA-129 with SNHG12 and SOX4. A series of rescue experiments were conducted to clarify the role of SNHG12/miRNA-129/SOX4 regulatory loop in the progression of OC. SNHG12 was upregulated in OC tissues relative to adjacent normal ones. Patients with metastatic OC or those in stage III-IV had a higher level of SNHG12 compared with non-metastatic or stage I-II patients. The 5-year survival was markedly worse in OC patients with high-level SNHG12 than those in the low-level group. Similarly, SNHG12 was highly expressed in OC cell lines. Overexpression of SNHG12 accelerated A2780 and HO8910 cells to proliferate and migrate. We observed the binding between SNHG12 and miRNA-129, and the latter was lowly expressed in OC. The miRNA-129 overexpression partially reversed the promotive effects of SNHG12 on proliferative and migratory abilities of OC cells. Subsequently, SOX4 was proved to be the target gene of miRNA-129. The SOX4 overexpression was further confirmed to reverse the inhibitory effects of miRNA-129 on proliferative and migratory abilities of OC cells. SNHG12 accelerates the proliferative and migratory abilities of OC cells via sponging miRNA-129 to upregulate SOX4.

MiR-27a promotes EMT in ovarian cancer through active Wnt/𝜷-catenin signalling by targeting FOXO1.

Ovarian cancer (OC) is the fifth most common type of cancer in women worldwide. MiR-27a plays an important role in the development of ovarian cancer. However, the exact function and molecular mechanism of miR-27a in epithelial-mesenchymal transition (EMT) has not been thoroughly elucidated to date. Quantitative real-time PCR (qRT-PCR) was used to determine the expression of miR-27a and FOXO1 mRNA in ovarian tissues and cells. The function of miR-27a in ovarian cancer was investigated through overexpression and knockdown of miR-27a in vitro. Wound healing and Transwell assays were performed to evaluate the migration and invasive capacity of the cells. A luciferase reporter assay was conducted to confirm the interaction between miR-27a and FOXO1. Western blotting was used to evaluate FOXO1, EMT and Wnt/β-catenin relative protein expression. In our study, we found that the mRNA expression level of miR-27a was significantly higher in ovarian cancer tissues and in HO8910 and OV90 cells. Functional experiments showed that miR-27a overexpression potentiated the migration and invasion of HO8910 and OV90 cells, while miR-27a inhibition reduced the cells' migration and invasion. Moreover, miR-27a upregulated the expression of mesenchymal cell markers and downregulated the expression of epithelial cell markers, which were restored via silencing of miR-27a expression. Subsequently, miR-27a was found to directly target and suppress the expression of FOXO1. Finally, we demonstrated that miR-27a promoted the progression of ovarian cancer cells and induced the process of EMT via the Wnt/β-catenin signalling pathway through inhibition of FOXO1. Taken together, these results indicate that targeting miR-27a and FOXO1 could represent a strategy for anticancer therapy in ovarian cancer.

Correlation of OPN gene expression with proliferation and apoptosis of ovarian cancer cells and prognosis of patients.

Correlation of osteopontin (OPN) gene expression with proliferation and apoptosis of ovarian cancer cells and prognosis of patients was investigated. The expression levels of OPN in 81 pairs of ovarian cancer tissues and para-carcinoma tissues obtained via surgical resection were detected using immunohistochemistry (IHC). The correlation of OPN protein expression with clinicopathological features of patients was analyzed. All patients were followed up for 3 years. The disease-free survival (DFS) and overall survival (OS) curves of patients in high/low OPN expression groups were drawn using the Kaplan-Meier method. The expression levels of OPN in normal ovarian epithelial IOSE80 cells and 5 ovarian cancer cell lines were detected via western blotting. Moreover, two cell lines with high OPN expression were interfered with lentiviral transfection technique. The effects of OPN on ovarian cancer cell proliferation and apoptosis were detected and analyzed via Cell Counting Kit-8 (CCK8) assay and flow cytometry. The positive expression rate of OPN protein in tumor tissues was higher than that in para-carcinoma tissues (P<0.05). Survival curves suggested that both DFS and OS in OPN negative group were superior to those in OPN positive group (P<0.05). Results of western blotting showed that OPN was weakly expressed in IOSE80 cells, whereas it was highly expressed in SKOV-3, COC1, A2780, HO-8910 and OVCAR-3 cells, among which the OPN protein expression levels were relatively higher in SKOV-3 and OVCAR-3 cell lines. After knockdown of OPN gene with sh-OPN, the cell proliferation rates of OVCAR-3 and SKOV-3 were significantly decreased from 48 h (P<0.05), but the apoptosis level was increased remarkably (28.2 vs. 1.3% and 25.3 vs. 3.2%), and differences were statistically significant (P<0.05). In conclusion, overexpression of OPN enhances the proliferation of ovarian cancer cells, which is an adverse factor for patient survival and prognosis.

MARCH2对卵巢癌HO-8910细胞自噬及生长的影响

MARCH2对卵巢癌HO-8910细胞自噬及生长的影响

LncRNA GIHCG promotes development of ovarian cancer by regulating microRNA-429.

To explore whether lncRNA GIHCG participates in the pathogenic progression of ovarian cancer (OC) and its underlying mechanism. Expression levels of GIHCG and microRNA-429 in 30 OC tissues and normal ovarian tissues were detected by quantitative Real time-polymerase chain reaction (qRT-PCR). Subsequently, 15 pairs of OC tissues and paracancerous tissues were selected for correlation analyses of GIHCG, microRNA-429 and the overall survival (OS) of OC patients using Kaplan-Meier method. Pearson correlation analyses were conducted for investigating the correlation between GIHCG and microRNA-429. GIHCG expression in OC cell lines (HEY, A2780 and HO8910) and normal epithelial OC cell line (IOSE-386) was detected by qRT-PCR. After transfection of GIHCG overexpression plasmid in HEY cells, cell cycle, proliferation and colony formation ability were detected by flow cytometry, cell counting kit-8 (CCK-8) and colony formation assay. MicroRNA-429 expression in HEY cells overexpressing GIHCG was detected by qRT-PCR. Rescue experiments were conducted by co-transfection of GIHCG overexpression plasmid and microRNA-429 mimics, followed by cell cycle and colony formation detection. GIHCG was highly expressed, whereas microRNA-429 was lowly expressed in OC tissues than that of paracancerous tissues. OC patients with higher expression of GIHCG showed shorter OS than those with lower expression. However, OC patients with higher expression of microRNA-429 had longer OS than those with lower expression. GIHCG expression was positively correlated to microRNA-429. In vitro experiments showed that GIHCG was highly expressed in HEY, A2780 and HO8910 cells than that of IOSE-386 cells. GIHCG overexpression in HEY cells promoted cell cycle and colony formation abilities, which were reversed by microRNA-429 overexpression. GIHCG is highly expressed in OC, which promotes OC development by stimulating cell cycle progression and cell proliferation by regulating microRNA-429.