微小RNA-6833-3p负调控LPIN1基因表达对肺癌细胞周期和增殖的影响  

Effect of microRNA-6833-3p on lung cancer cell cycle and proliferation through negative regulation of LPIN1 gene expression

作　　者：张静宜 夏蕾 张晓月 谭本旭 杨镇洲 ZHANG Jingyi;XIA Lei;ZHANG Xiaoyue;TAN Benxu;YANG Zhenzhou(Cancer Center,Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China)

机构地区：[1]重庆医科大学附属第二医院肿瘤中心,重庆400010

出　　处：《山西医科大学学报》2021年第7期807-811,共5页Journal of Shanxi Medical University

基　　金：国家自然科学基金面上项目(81972851)。

摘　　要：目的探讨微小RNA-6833-3p(miR-6833-3p)通过靶向调控脂蛋白1(lipin 1,LPIN1)的表达影响肺癌细胞周期和增殖的作用及机制。方法采用实时定量PCR(qPCR)检测miR-6833-3p在肺癌细胞(HCC827、A549、H1650、H1299)和永生化正常肺泡上皮细胞(HPAEpiC)中的表达水平。以携带miR-6833-3p的慢病毒或阴性对照慢病毒感染表达最低的肺癌细胞,分别命名为miR-6833-3p组和对照组。流式细胞术和四甲基偶氮唑蓝(MTT)法分别检测miR-6833-3p对肺癌细胞周期和增殖的影响。采用生物信息学软件预测miR-6833-3p的潜在靶基因,采用双荧光素酶报告基因实验进行验证miR-6833-3p和靶基因的结合。qPCR和Western blot检测miR-6833-3p对靶基因和细胞周期蛋白表达的影响。结果与HPAEpiC相比,miR-6833-3p在肺癌细胞中均呈现低表达(P<0.05),H1299细胞中的表达最低(P<0.01)。与对照组比较,miR-6833-3p组H1299细胞中miR-6833-3p的表达水平明显增加(P<0.01)。与对照组相比,miR-6833-3p组中G0/G1期细胞比例明显增加(P<0.01),S期和G2/M期细胞比例明显减少(P<0.01)。与对照组比较,miR-6833-3p组H1299细胞的增殖从第3天开始明显降低(P<0.05)。生物信息学软件预测miR-6833-3p的潜在靶基因是LPIN1,miR-6833-3p可与LPIN1 mRNA 3′非翻译区靶向结合(P<0.01)。与对照组相比,miR-6833-3p组LPIN1基因表达显著减少(P<0.01),细胞周期蛋白CDK2、CDK4及Cyclin D1表达显著降低(P<0.01)。结论miR-6833-3p在肺癌细胞中表达降低,miR-6833-3p通过干扰LPIN1基因的表达,抑制肺癌H1299细胞周期和增殖,miR-6833-3p具有成为肺癌诊疗新靶点的潜力。Objective To explore the effect of microRNA-6833-3p(miR-6833-3p)on lung cancer cell cycle and proliferation through targeted regulation of lipin 1(LPIN1)expression and its mechanism.Methods Real-time quantitative PCR(qPCR)was used to detect the expression level of miR-6833-3p in lung cancer cells(HCC827,A549,H1650,H1299)and immortalized normal alveolar epithelial cell lines(HPAEpiC).The lung cancer cells with the lowest expression were infected with the lentivirus carrying miR-6833-3p or the negative control lentivirus,and named as miR-6833-3p group and control group.The effects of miR-6833-3p on lung cancer cell cycle and proliferation were detected by flow cytometry and tetramethylazazole blue(MTT)method,respectively.Bioinformatics software was used to predict the potential target gene of miR-6833-3p,and the dual luciferase reporter gene experiment was used to verify the binding of miR-6833-3p to the target gene.QPCR and Western blot were used to detect the effect of miR-6833-3p on the expression of target gene and cell cycle-related proteins.Results Compared with HPAEpiC cells,the expression of miR-6833-3p was decreased in lung cancer cells(P<0.05),and the expression in H1299 cells was the lowest(P<0.01).Compared with control group,the expression level of miR-6833-3p in H1299 cells was significantly increased in miR-6833-3p group(P<0.01).Compared with control group,the percentage of G0/G1 phase cells was significantly increased in miR-6833-3p group(P<0.01),while the percentage of S phase and G2/M phase cells was significantly reduced(P<0.01).Compared with control group,the proliferation of H1299 cells in miR-6833-3p group was significantly reduced from day 3(P<0.05).Bioinformatics software predicted that the potential target gene of miR-6833-3p was LPIN1.The miR-6833-3p could targetedly bind to the 3′untranslated region of LPIN1 mRNA(P<0.01).Compared with control group,the expression of LPIN1 gene in miR-6833-3p group was significantly reduced(P<0.01),and the expression of CDK2,CDK4 and Cyclin D1 was signifi

关 键 词：miR-6833-3p 肺癌 细胞周期 细胞增殖 

分 类 号：R734.2[医药卫生—肿瘤]