MiR-152-3p靶向PIK3CA基因对乳腺癌细胞生物学特性的调控及其机制  

Regulation and mechanism of miR-152-3p targeting PIK3CA gene on biological characteristics of breast cancer cells

作　　者：潘蕾蕾 江卫兵 方芳[2] 陈剑平[2] 殷照才[2] PAN Lei-lei;JIANG Wei-bing;FANG Fang;CHEN Jian-ping;YIN Zhao-cai(Department of Cardiothoracic and Breast Surgery,Xuancheng Central Hospital,Xuancheng 242000,Anhui Province,China;Department of Thyroid and Breast Surgery,Yijishan Hospital of Wannan Medical College,Wannan 241001,Anhui Province,China)

机构地区：[1]宣城市中心医院心胸乳腺外科,安徽宣城242000 [2]皖南医学院弋矶山医院甲状腺乳腺外科,安徽皖南241001

出　　处：《新乡医学院学报》2019年第12期1130-1136,共7页Journal of Xinxiang Medical University

摘　　要：目的探讨miR-152-3p靶向PIK3CA基因对乳腺癌细胞生物学特性的调控及机制。方法选取2016年1月至2017年7月宣城市中心医院保存的乳腺癌组织和相应的正常乳腺组织(距肿瘤边缘>5 cm)标本各50例,培养乳腺癌MCF-7细胞和正常乳腺上皮细胞MCF-10A,采用反转录定量聚合酶链反应(qRT-PCR)检测乳腺癌组织、正常乳腺组织、MCF-7细胞及MCF-10A细胞中miR-152-3p及PIK3CA mRNA表达。培养MCF-7细胞,待细胞融合度达30%~50%时将细胞分为空白组、阴性对照组、miR-152-3p mimic组、miR-152-3p inhibitor组、si-PIK3CA组和miR-152-3p inhibitor+si-PIK3CA组,空白组细胞不转染任何序列,阴性对照组细胞转染miR-152-3p阴性对照质粒,miR-152-3p mimic组细胞转染miR-152-3p mimic质粒,miR-152-3p inhibitor组细胞转染miR-152-3p inhibitor质粒,si-PIK3CA组细胞转染si-PIK3CA质粒,miR-152-3p inhibitor+si-PIK3CA组细胞共转染miR-152-3p inhibitor质粒和si-PIK3CA,转然后继续培养24~48 h;采用Western blot法检测各组MCF-7细胞中PIK3CA、细胞外信号调节激酶(ERK)、c-Jun氨基末端激酶(JNK)、p38和E-cadherin蛋白表达,四甲基偶氮唑盐法检测各组MCF-7细胞增殖活性,划痕实验检测各组MCF-7细胞迁移能力,Transwell侵袭实验检测各组MCF-7细胞侵袭能力。结果乳腺癌组织中miR-152-3p相对表达量显著低于正常乳腺组织,PIK3CA mRNA相对表达量显著高于正常乳腺组织(P<0.05)。MCF-7细胞中miR-152-3p相对表达量显著低于MCF-10A细胞,PIK3CA mRNA相对表达量显著高于MCF-10A细胞(P<0.05)。与空白组和阴性对照组比较,miR-152-3p mimic组和si-PIK3CA组MCF-7细胞中PIK3CA、p38、ERK、JNK和E-cadherin蛋白相对表达量显著降低,miR-152-3p inhibitor组MCF-7细胞中PIK3CA、p38、ERK、JNK和E-cadherin蛋白相对表达量显著升高(P<0.05)。miR-152-3p inhibitor+si-PIK3CA组与空白组和阴性对照组MCF-7细胞中PIK3CA、p38、ERK、JNK和E-cadherin蛋白相对表达量比较差异无统计学意义(Objective To investigate the regulation and mechanism of miR-152-3p targeting PIK3CA gene on biological characteristics of breast cancer cells.Methods Fifty samples of breast cancer tissues and 50 samples of corresponding normal breast tissues(more than 5 cm from the edge of the tumor)were collected in Xuancheng Central Hospital from January 2016 to July 2017.Breast cancer MCF-7 cells and normal breast epithelium MCF-10A cells were cultured.The mRNA expression of miR-152-3p and PIK3CA in breast cancer tissues,normal breast tissues,MCF-7 cells and MCF-10A cells were detected by reverse transcription quantitative polymerase chain reaction(qRT-PCR).When the fusion degree of MCF-7 cells reached 30%-50%,the cells were divided into blank group,negative control group,miR-152-3p mimic group,miR-152-3p inhibitor group,si-PIK3CA group and miR-152-3p inhibitor+si-PIK3CA group.The cells in the blank group were not transfected with any sequence;the cells in the negative control group,miR-152-3p mimic group,miR-152-3p inhibitor group and si-PIK3CA group were transfected with miR-152-3p negative control plasmids,miR-152-3p mimic plasmids,miR-152-3p inhibitor plasmids and si-PIK3CA plasmids,respectively;the cells in the miR-152-3p inhibitor+si-PIK3CA group were transfected with miR-152-3p inhibitor plasmids and si-PIK3CA.Then the cells were cultured for 24-48 hours.The expressions of PIK3CA,extracellular signal-regulated kinase(ERK),Jun N-terminal kinase(JNK),p38 and E-cadherin protein in MCF-7 cells were detected by Western blot.The proliferation activity of MCF-7 cells was detected by methyl thiazolyl tetrazolium method.The migration ability of MCF-7 cells was detected by scratch test,and the invasion ability of MCF-7 cells in each group was detected by Transwell invasion test.Results The relative expression of miR-152-3p in the breast cancer tissues was significantly lower than that in the normal breast tissues,and the relative expression of PIK3CA mRNA in the breast cancer tissues was significantly higher than that in the no

关 键 词：乳腺癌 miR-152-3p PIK3CA 丝裂原活化蛋白激酶信号通路 细胞增殖 

分 类 号：R737.9[医药卫生—肿瘤]