miR-126靶向SETD8抑制乳腺癌细胞增殖、迁移的作用及其机制研究  

Inhibitory effect of miR-126 targeting SETD8 on proliferation and migration of breast cancer cells and its mechanism

作　　者：张晓静[1] 陈延松[1] 陈晨[1] 刘元[1] 金功圣[1] 刘先富[1] ZHANG Xiaojing;CHEN Yansong;CHEN Chen;LIU Yuan;JIN Gongsheng;LIU Xianfu(Department of Surgical Oncology,First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠医学院第一附属医院肿瘤外科,蚌埠233000

出　　处：《山西医科大学学报》2020年第12期1308-1314,共7页Journal of Shanxi Medical University

基　　金：安徽省教育厅自然科学项目(KJ2019A0341,KJ2017A245);蚌埠市科技创新指导类项目(20190309)。

摘　　要：目的探究miR-126对乳腺癌MCF-7细胞增殖、迁移的影响及其分子机制。方法提取乳腺上皮MCF-10A细胞和乳腺癌MDA-MB-231、MCF-7、SK-BR-3、BT-549细胞总RNA,使用qRT-PCR检测miR-126的表达水平。将MCF-7细胞分为阴性对照组(mimics NC)和miR-126模拟物转染组(miR-126 mimics),CCK-8和集落克隆实验检测细胞增殖,Transwell实验检测细胞迁移。Targetscan网站在线预测miR-126下游靶基因,Western blot检测miR-126对SETD8和PCNA表达的影响;挽救实验探究SETD8表达对细胞增殖、迁移能力的影响。结果相比于乳腺上皮细胞MCF-10A,miR-126在四株乳腺癌细胞中均降低(P<0.05)。与mimics NC组比较,miR-126 mimics组细胞增殖能力降低、集落克隆数目减少、细胞迁移数目降低(P<0.05)。Western blot结果证实miR-126能抑制SETD8蛋白表达,且miR-126能降低PCNA蛋白表达。挽救实验证实,相比于miR-126 mimics组,SETD8回补组能够部分恢复乳腺癌细胞增殖能力、增加集落克隆形成和细胞迁移数目(P<0.05),并能增加PCNA蛋白表达。结论miR-126能抑制乳腺癌细胞增殖和迁移,miR-126靶向SETD8抑制PCNA表达可能是其关键的分子机制。Objective To explore the effect of miR-126 on the proliferation and the metastasis of breast cancer MCF-7 cells and its molecular mechanism.Methods The total RNA of breast epithelial MCF-10A cells and breast cancer MDA-MB-231,MCF-7,SK-BR-3,BT-549 cells was extracted,and the expression level of miR-126 was detected by qRT-PCR.MCF-7 cells were divided into negative control group(mimics NC)and miR-126 mimics transfection group(miR-126 mimics),CCK-8 and colony cloning experiments were used to detect the cell proliferation,and Transwell experiments were used to detect the cell migration.The downstream target gene of miR-126 was predicted online through the Targetscan website.Western blot was used to detect the effect of miR-126 on the expression of SETD8 and PCNA.Rescue experiment was used to explore the effect of SETD8 expression on cell proliferation and metastasis.Results Compared with breast epithelial cells MCF-10A,miR-126 was reduced in all four breast cancer cells(P<0.05).Compared with mimics NC group,the cell proliferation ability,the number of clones and the number of cell migration were reduced in miR-126 mimics group(P<0.05).Western blot results confirmed that miR-126 inhibited SETD8 protein expression,and miR-126 also reduced PCNA protein expression.The rescue experiment confirmed that compared with miR-126 mimics group,the breast cancer cell proliferation was partially restored in SETD8 supplementation group,the number of colony formation and the cell migration were increased(P<0.05),and PCNA protein expression was increased.Conclusion The miR-126 may inhibit the breast cancer cell proliferation and the migration by suppressing PCNA expression via targeting SETD8.

关 键 词：乳腺癌 MIR-126 SETD8 PCNA 增殖 迁移 

分 类 号：R737.9[医药卫生—肿瘤]