miR-493-5p抑制人卵巢癌细胞系SKOV3增殖和迁移

miR-493-5p inhibits the proliferation and migration of human ovarian cancer cell line SKOV3

目的探讨miR-493-5p对卵巢癌细胞的增殖、侵袭和迁移的影响以及潜在的作用机制。方法培养人卵巢上皮细胞系IOSE80以及卵巢癌细胞系A2780、SKOV3和OVCAR3,将miR-493-5p过表达载体、miR-493-5p抑制表达载体及G蛋白偶联受体激酶相互作用因子1(GIT1)表达载体转染至SKOV3细胞,采用RT-qPCR检测miR-493-5p和mRNA的表达水平;Western blot检测蛋白表达;MTT法检测细胞活性;Transwell小室法检测细胞迁移和侵袭;双荧光素酶报告实验检测miR-493-5p和GIT1的靶向关系。结果A2780、SKOV3和OVCAR3细胞中miR-493-5p低表达,GIT1高表达(P<0.05);过表达miR-493-5p后,SKOV3细胞增殖活性、迁移和侵袭数降低(P<0.05)。miR-493-5p靶向调控GIT1的表达,过表达GIT1降低了miR-493-5p对SKOV3细胞增殖、迁移和侵袭的抑制作用以及对p-MEK1/2、p-ERK1/2、p-c-Jun、p-c-Fos表达的抑制作用。结论miR-493-5p可抑制卵巢癌SKOV3细胞的增殖、迁移和侵袭,其机制可能与靶向调控GIT1以及MEK1/2-ERK1/2通路有关,并可为卵巢癌的诊断、预防和治疗提供新靶点。

Objective To investigate the effects of miR-493-5p on proliferation,invasion and migration of ovarian cancer cells and underlying mechanism.Methods Culture human normal ovarian epithelial cell lines IOSE80 and ovarian cancer cell lines A2780,SKOV3,OVCAR3,transfected the miR-493-5p over-expression vectors,miR-493-5p suppressed expression vectors and G protein coupled receptor kinase interacting factor 1(GIT1)over-expression vectors into SKOV3 cells;RT-qPCR was used to detect miR-493-5p and GIT1 mRNA expression;Western blot was used to detect protein expression;MTT assay was used to detect cell viability;Transwell was used to detect cell migration and invasion;dual luciferase report experiment was applied for detection of the targeting relationship between miR-493-5p and GIT1.Results In ovarian cancer cells A2780,SKOV3,OVCAR3,miR-493-5p were weakly expressed,and GIT1 was highly expressed(P<0.05);After over-expression of miR-493-5p,the proliferation activity,migration and invasion number of ovarian cancer cell SKOV3 were decreased(P<0.05).miR-493-5p targeted at the regulation of GIT1 expression,and over-expression of GIT1 reduced the inhibitory effect of miR-493-5p on proliferation,migration and invasion of ovarian cancer cell SKOV3 and expression of p-MEK1/2,p-ERK1/2,pc-Jun and pc-Fos.Conclusions miR-493-5p can inhibit the proliferation,migration and invasion of ovarian cancer cells SKOV3.The mechanism may be related to the targeted regulation of GIT1 and MEK1/2-ERK1/2 pathway,which thus provides some new targets for diagnosis,prevention and treatment for ovarian cancer.