LncRNA OIP5-AS1对卵巢上皮IOSE80表型转化的影响

Effect of LncRNA OIP5-AS1 on Phenotypic Transformation of IOSE80 in Ovarian Epithelium

目的探索lncRNA OIP5-AS1对卵巢上皮细胞IOSE80发生增殖、迁移、凋亡、侵袭和周期的表型的变化及可能的机制。方法患者临床资料数据收集自TCGA数据库及GEO数据库,R包分析后可视化OIP5-AS1在卵巢癌组织中表达上调,生存率与OIP5-AS1的相关性采用Kaplan-Meier分析。以慢病毒载体构建OIP5-AS1过表达和沉默的IOSE80细胞模型,采用RT-qPCR验证OIP5-AS1的表达,CCK-8检测细胞增殖,Transwell检测侵袭,划痕试验检测细胞迁移,流式细胞术检测细胞周期和凋亡,Western blotting检测细胞侵袭相关蛋白E钙黏着蛋白(E-cadherin)和N钙黏着蛋白(Ncadherin)的表达及细胞周期蛋白依赖性激酶(cyclin-dependent kinase,CDK)和细胞周期蛋白G相关激酶(cyclin-G-related kinase,GAK)的表达。结果RT-qPCR结果显示成功构建OIP5-AS1过表达和沉默的IOSE80细胞株。CCK-8结果显示过表达OIP5-AS1促进IOSE80细胞的增殖。划痕试验结果显示过表达OIP5-AS1促进IOSE80细胞迁移。Transwell结果显示过表达OIP5-AS1会引起IOSE80细胞的侵袭力增强。流式细胞术结果表明OIP5-AS1的过表达使IOSE80细胞凋亡减弱并推动了细胞周期的进展。Western blotting结果显示过表达OIP5-AS1会下调E-cadherin的表达并上调N-cadherin的表达,同时过表达OIP5-AS1可提高CDK及GAK蛋白的表达。结论lncRNA OIP5-AS1通过上调CDK及GAK的表达进一步干预了IOSE80细胞周期的调控,进而实现对卵巢上皮细胞恶性表型的间接调控作用。

OBJECTIVE To explore the phenotypic changes and possible mechanisms of lncRNA OIP5-AS1 on the proliferation,migration,apoptosis,invasion and cycle of ovarian epithelial cells IOSE80.METHODS The clinical data of patients were collected from TCGA database and GEO database.After R package analysis,the differential expression of OIP5-AS1 was visualized in the volcanic map.The correlation between survival rate and OIP5-AS1 was analyzed by Kaplan-Meier.The IOSE80 cell model of OIP5-AS1 over expression and silencing was constructed with lentivirus vector.The expression of OIP5-AS1 was verified by RT-qPCR.Cell proliferation was detected by CCK-8.Invasion was detected by Transwell.Cell migration was detected by scratch test.Cell cycle and apoptosis were detected by flow cytometry.Western blotting was used to detect the expression of E-cadherin and N-cadherin,as well as the expression of cyclin-dependent kinase(CDK)and cyclin-G-related kinase(GAK).RESULTS RT-qPCR results showed that IOSE80 cell lines over expressing and silencing OIP5-AS1 were successfully constructed.CCK-8 results showed that overexpressing OIP5-AS1 promoted the proliferation of IOSE80 cells.Scratch test results showed that overexpressing OIP5-AS1 promoted the migration of IOSE80 cells.Transwell results showed that overexpressing OIP5-AS1 would increase the invasiveness of IOSE80 cells.Flow cytometry results showed that overexpression of OIP5-AS1 weakened the apoptosis of IOSE80 cells and promoted the progress of cell cycle.Western blotting results showed that overexpression of OIP5-AS1 downregulated the expression of E-cadherin and upregulated the expression of N-cadherin,while overexpression of OIP5-AS1 increased the expression of CDK and GAK proteins.CONCLUSION LncRNA OIP5-AS1 further interferes with the regulation of IOSE80 cell cycle by up regulating the expression of CDK and GAK,and then indirectly regulates the malignant phenotype of ovarian epithelial cells.