抑癌基因VHL与RNASET2甲基化协同microRNA调控卵巢癌细胞耐药逆转

Tumor Suppressor Gene VHL and RNASET2 Methylation Cooperate with microRNA to Regulate the Reversal of Drug Resistance in Ovarian Cancer Cells

目的 研究卵巢癌耐药细胞中抑癌基因甲基化和miRNA的关系以及对耐药逆转的影响。方法 基于基因表达公共数据库(GEO)筛选2个卵巢癌多药耐药相关数据集(GSE176218 Public on May 16,2022;GSE198077 Public on Apr 20,2022)鉴定抑癌基因。再通过甲基化位点分析,miRNA预测和GeneMANIA互作网络分析进行抑癌基因甲基化与miRNA的关系探讨。MTT和RT-PCR检测DNA甲基化抑制剂和(或)miRNA抑制剂作用下抑癌基因对耐药的影响。结果 卵巢癌耐药细胞中VHL和RNASET2表达下调,且与患者预后相关。VHL和RNASET2受到DNA甲基化调控。hsa-miR-635靶向RNASET2,hsa-miR-106b靶向VHL。VHL、RNASET2甲基化和miRNA协同调控。VHL与RNASET2重新表达通过PI3K/AKT信号通路逆转耐药。结论DNA甲基化和miRNA协同调控抑癌基因RNASET2和VHL的表达,并逆转卵巢癌细胞的多药耐药。

Objective To study the relationship between tumor suppressor gene methylation and miRNA in drug-resistant ovarian cancer cells and its effect on drug resistance reversal.Methods Multidrug resistance tumor suppressor genes of ovarian cancer were screened based on GEO database.Then,the relationship between tumor suppressor gene methylation and miRNA was explored through methylation site analysis,miRNA prediction and GeneMANIA interaction network analysis.MTT and RT-PCR were used to detect the effect of tumor suppressor genes on drug resistance under the action of DNA methylation inhibitors and/or miRNA inhibitors.Results The expressions of VHL and RNASET2 in ovarian cancer drug-resistant cells were down-regulated and correlated with the prognosis of patients.VHL and RNASET2 are regulated by DNA methylation.hsa-miR-635 targets RNASET2 and hsa-miR-106b targets VHL.VHL,RNASET2 methylation and miRNA co-regulation.VHL and RNASET2 re-expression reversed drug resistance through PI3K/AKT signaling pathway.Conclusions DNA methylation and miRNA synergistically regulate the expression of tumor suppressor genes RNASET2 and VHL,and reverse the multidrug resistance of ovarian cancer cells.