miR-3911对卵巢癌细胞增殖和迁移的影响及其机制

Effect of miR-3911 on proliferation and migration of ovarian cancer cells and its mechanism

目的探讨微小RNA(miRNA)-3911对卵巢癌细胞增殖和迁移能力的影响及其作用机制。方法实时定量聚合酶链式反应(qRT-PCR)检测卵巢癌细胞株(A2780、OC3、HO-8910、SKOV-3)及正常卵巢上皮细胞(IOSE80)中miR-3911的表达水平,选择miR-3911表达水平最低的卵巢癌细胞作为实验对象,分为两组:control组和miR-3911组,分别转染无意义序列NC mimic和miR-3911 mimic。qRT-PCR检测转染细胞中miR-3911的表达水平。采用CCK-8法和Transwell迁移实验分别检测miR-3911对卵巢癌细胞增殖和迁移能力的影响。通过靶基因预测网站microRA.org、PITA和miRTarBase预测miR-3911的靶基因,并采用双荧光素酶报告基因实验验证。qRT-PCR和Western blot检测转染细胞中miR-3911靶基因的表达水平。Western blot检测NF-κB信号通路蛋白NF-κB、p-IκBα、p-p65表达。结果与IOSE80细胞相比,卵巢癌细胞株(A2780、OC3、HO-8910、SKOV-3)中miR-3911的表达水平显著降低(P<0.01),HO-8910细胞中miR-3911表达水平最低(P<0.01)。与control组相比,miR-3911组HO-8910细胞增殖能力明显降低(P<0.05),迁移能力受到抑制(P<0.01)。靶基因预测网站和双荧光素酶报告基因结果显示,NF-κB激活蛋白(NF-κB activating protein,NKAP)是miR-3911的靶基因(P<0.01)。与control组比较,miR-3911组HO-8910细胞中NKAP基因表达水平显著降低(P<0.01),NF-κB信号通路蛋白NF-κB、p-IκBα、p-p65表达降低(P<0.01)。结论miR-3911在卵巢癌细胞株中呈低表达,miR-3911通过靶向负调控NKAP表达抑制NF-κB信号通路活化,降低卵巢癌HO-8910细胞的增殖和迁移能力。

Objective To explore the effect of microRNA(miRNA)-3911 on the proliferation and migration of ovarian cancer cells and its mechanism.Methods Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression level of miR-3911 in ovarian cancer cell lines A2780,OC3,HO-8910,SKOV-3 and normal ovarian epithelial cells IOSE80.Ovarian cancer cells with the lowest expression level of miR-3911 were used as experimental subjects and were divided into two groups:control group(transfected with nonsense sequence NC mimic)and miR-3911 group(transfected with miR-3911 mimic).RT-qPCR was used to detect the expression level of miR-3911 in transfected cells.CCK-8 method and Transwell migration test were used to detect the effects of miR-3911 on the proliferation and migration of ovarian cancer cells.The target gene prediction websites microRA.org,PITA and miRTarBase were used to predict the target gene of miR-3911,and the dual luciferase reporter gene experiment was used to verify it.RT-qPCR and Western blot were used to detect the expression level of miR-3911 target gene in transfected cells.Western blot was used to detect the expression of NF-κB signaling pathway proteins NF-κB,p-IκBα,and p-p65.Results Compared with IOSE80 cells,the expression level of miR-3911 in ovarian cancer cell lines A2780,OC3,HO-8910,SKOV-3 was significantly reduced(P<0.01),and it was the lowest in HO-8910 cells(P<0.01).Compared with control group,the proliferation ability of HO-8910 cells was significantly reduced in miR-3911 group(P<0.05),and the migration ability was inhibited(P<0.01).The target gene prediction websites and dual luciferase reporter gene results showed that NF-κB activating protein(NKAP)was the target gene of miR-3911(P<0.01).Compared with control group,the expression level of NKAP gene in HO-8910 cells was significantly reduced in miR-3911 group(P<0.01),and the expression of NF-κB signaling pathway proteins NF-κB,p-IκBα,and p-p65 was decreased(P<0.01).Conclusion The miR-3911 is lowly expressed in ovarian cancer cell lines,and miR-3911 can inhibit the activation of NF-κB signaling pathway by negatively targetedly regulating the expression of NKAP,and reduce the proliferation and migration abilities of ovarian cancer HO-8910 cells.