miRNA-186-3p对子宫颈癌CaSki细胞功能影响及与转化生长因子β-Smad信号通路关系的研究  

Effect of miRNA-186-3p on the function of cervical cancer CaSki cells and its relationship with transforming growth factorβ-Smad signaling pathway

作　　者：王敬苗 李静[2] 李晓丹[3] 赵航 Wang Jingmiao;Li Jing;Li Xiaodan;Zhao Hang(Department of Outpatient,Handan First Hospital,Handan 056002,China;Department of Oncology 3,Handan Central Hospital,Handan 056001,China;Department of Gynecology,Handan First Hospital,Handan 056002,China)

机构地区：[1]邯郸市第一医院门诊部,邯郸056002 [2]邯郸市中心医院肿瘤三科,邯郸056001 [3]邯郸市第一医院妇科,邯郸056002

出　　处：《肿瘤研究与临床》2022年第10期737-740,共4页Cancer Research and Clinic

基　　金：河北省医学科学研究重点课题(20181640)。

摘　　要：目的探讨miRNA-186-3p(miR-186-3p)对子宫颈癌CaSki细胞凋亡、迁移、侵袭的影响及与转化生长因子β(TGF-β)-Smad信号通路的关系。方法将载有miR-186-3p抑制基因、miR-186-3p模拟物的质粒分别转染至子宫颈癌CaSki细胞,分别为miR-186-3p-I组、miR-186-3p-M组;另将转染空载质粒的CaSki细胞作为对照,为miR-186-3p-C组。采用流式细胞术检测各组细胞凋亡率,采用Transwell法检测各组细胞迁移和侵袭能力;采用蛋白质印迹法检测各组细胞TGF-β-Smad信号通路相关蛋白表达水平;采用Starbase软件预测miR-186-3p靶基因,并用双荧光素酶报告基因实验进行验证。结果miR-186-3p-M组细胞凋亡率均低于miR-186-3p-I组和miR-186-3p-C组[(7.5±3.2)%比(13.9±0.7)%、(12.7±0.6)%,均P<0.05]。miR-186-3p-M组迁移细胞数[(218±25)个比(168±13)个、(175±13)个,均P<0.001]和侵袭细胞数均多于miR-186-3p-I组和miR-186-3p-C组[(165±21)个比(130±11)个、(142±12)个,均P<0.001]。miR-186-3p-M组TGF-β、Smad2、Smad3、磷酸化Smad2(p-Smad2)、磷酸化Smad3(p-Smad3)蛋白相对表达量均最低,与另两组比较,差异均有统计学意义(均P<0.001)。采用Starbase软件预测miR-186-3p与XIST RNA互补结合;双荧光素酶报告基因实验显示,XIST野生型载体+miR-186-3p模拟物质粒共转染的细胞相对荧光素酶活性低于XIST野生型载体+miR-186-3p空载质粒共转染的细胞(P<0.001)。结论miR-186-3p可能直接与XIST RNA结合而下调TGF-β-Smad信号通路活性,进而增强子宫颈癌CaSki细胞凋亡、迁移、侵袭活性。Objective To investigate the effect of miRNA-186-3p(miR-186-3p)on the apoptosis,immigration and invasion of cervical cancer CaSki cells and its relationship with transforming growth factor-β(TGF-β)-Smad signaling pathway.Methods Plasmids containing miR-186-3p suppressor gene and miR-186-3p mimics were transfected into cervical cancer CaSki cells,namely miR-186-3p-I group and miR-186-3p-M group,respectively.In addition,CaSki cells transfected with empty plasmids were used as control(miR-186-3p-group C).Flow cytometry was used to detect the apoptosis rate of each group,and Transwell method was used to detect the migration and invasion ability of each group.Western blotting was used to detect the expression level of TGF-β-Smad signaling pathway related proteins.The target genes of miR-186-3p were predicted by using Starbase software and verified by using dual luciferase reporter gene assay.Results The apoptosis rate of miR-186-3p-M group was lower than that of miR-186-3p-I group and miR-186-3p-C group[(7.5±3.2)%vs.(13.9±0.7)%,(12.7±0.6)%,all P<0.05].The number of migrating cells in miR-186-3p-M group[(218±25)vs.(168±13),(175±13),both P<0.001]and the number of invasive cells in miR-186-3p-I group and miR-186-3p-C group were higher than those in miR-186-3p-I group and miR-186-3p-C group[(165±21)vs.(130±11),(142±12),all P<0.001].The relative expression levels of TGF-β,Smad2,Smad3,phosphorylated Smad2(p-Smad2)and phosphorylated Smad3(p-Smad3)in miR-186-3p-M group were the lowest,and the differences were statistically significant compared with the other two groups(all P<0.001).Starbase software was used to predict the complementary binding of miR-186-3p to XIST RNA.Dual luciferase reporter assay showed that the relative luciferase activity of cells co-transfected with XIST wild-type vector and miR-186-3p mimic plasmid was lower than that of cells co-transfected with XIST wild-type vector and miR-186-3p empty plasmid(P<0.001).Conclusion miR-186-3p may directly bind to XIST RNA and down-regulate the activity o

关 键 词：子宫肿瘤 转化生长因子Β SMAD蛋白质类 miRNA-186-3p 细胞增殖 细胞凋亡 肿瘤侵润 

分 类 号：R737.33[医药卫生—肿瘤]