miR-19a在结肠癌中高表达以及通过靶定DLC1促进结肠癌细胞增殖  

Effect of high expressed miR-19a,on the proliferation of colon cancer cells through the target gene of DLC1

作　　者：郭运生[1,2] 戚峰[1] 刘彤[1] 王益民[2] 周文阔[2] 

机构地区：[1]天津医科大学总医院普通外科普通外科研究所,天津300052 [2]秦皇岛市第一医院普外二科,秦皇岛066000

出　　处：《天津医科大学学报》2013年第3期182-185,共4页Journal of Tianjin Medical University

摘　　要：目的:探讨miR-19a在结肠癌组织中的表达对结肠癌细胞系SW620和SW480增殖的影响,以及其靶基因的确定和功能性研究。方法:实时荧光定量PCR检测miR-19a的表达。噻唑蓝(MTT)比色实验和克隆形成实验检测SW620和SW480细胞的增殖活性。生物信息学预测、荧光报告载体实验以及实时荧光定量PCR和western blot验证miR-19a下游靶基因。结果:miR-19a在结肠癌组织中表达增强(P<0.01)。miR-19a可以增强结肠癌细胞系SW620和SW480增殖能力。DLC1是miR-19a的候选靶基因,过表达miR-19a可以同时在mRNA和蛋白水平上抑制DLC1基因的表达(P<0.01),反之抑制miR-19a的表达后DLC1的表达则升高(P<0.01)。过表达DLC1后,SW620和SW480细胞的活性和克隆形成能力减弱(P<0.01)。结论:DLC1是miR-19a的直接靶基因,miR-19a通过抑制DLC1的表达而促进结肠癌细胞的增殖。Objective： To investigate the expression level of miR-19a in colon carcinoma tissues; the role of miR-19a on the colon can- cer cell proliferation and identify the target gene for miR-19a in colon cancer cells. Methods： The expression of miR-19a were detected by real time PCR. The proliferation ability of the transfected colon cancer cells were measured by MTT and colony formation assay meth- ods. The bioinforamtics was used to predict the target gene for miR-19a, and EGFP reporter assay, real time PCR and western blot were performed to validate the target gene. Results： miR-19a was up-regulated in the colon carcinoma tissues, compared to the adjacent non- tumor tissues （P〈0.01）. miR-19a enhanced the cell viability and colony formation （P〈0.01）, in contrast, inhibition of miR-19a with miR- 19a ASO had the opposite effect （P〈0.01）. DLCI was predicted as a candidate target gene for miR-19a, miR-19a reduced the DLC1 ex- pression on the mRNA and protein levels （P〈0.01）, in contrast, inhibition of miR-19a had the opposite effect. Finally, the overexpression of DLC1 reduced the colon cancer cells viability and colony formation ability （P〈0.01）. Conclusion： DLC is a direct target gene for miR- 19a, and miR-19a promots the cell proliferation by downregulation of DLC1.

关 键 词：微小RNA 结肠癌 增殖 DLC1 

分 类 号：R735.35[医药卫生—肿瘤]