摘要
目的检测MicroRNA-128a(miR-128a)在肝细胞癌中的表达情况并探讨其在肝癌细胞中的生物学功能及机制。方法收集19例肝细胞癌及对应癌旁组织,运用qRT-PCR技术检测miR-128a在肝细胞癌和对应癌旁组织及肝癌细胞株中的表达;应用miR-128a mimics或inhibitor转染人肝癌细胞,MTT检测转染后细胞增殖活力变化;软件预测miR-128a的潜在靶点RND3,并用双荧光素酶报告实验证实miR-128a与RND3 3'UTR结合,qRT-PCR和Western blot检测miR-128a潜在靶点RND3的表达变化;Western blot检测细胞周期蛋白变化。结果在肝细胞癌组织中miR-128a的表达显著高于对应的癌旁组织(P<0.05);miR-128a促进肝癌细胞增殖(P<0.05);在肝癌细胞中miR-128a通过与RND3 3'URT结合下调RND3 mRNA和蛋白的表达(P<0.05);抑制miR-128a的表达可以导致cyclin B1、cyclin D1和CDK4表达下调。结论 miR-128a在肝细胞癌中表达上调并通过靶向调控RND3促进肝癌细胞增殖。
Objective To investigate the expression of microRNA- 128a（miR- 128a） and its role in hepatocellular carcinoma（HCC）. Methods Nineteen pairs of fresh surgical specimens of HCC and adjacent tissues were examined for miR- 128 a expression using qRT-PCR. A miR-128 a mimics or inhibitor was transfected into HCC cells, and the cell viability was analyzed by MTT assay. RND3, one of the potential targets of miR-128 a, was predicted by bioinformatics software and demonstrated by dual luciferase reporter assay. The expression of RND3 after transfection was detected using qRT-PCR and Western blotting,and the cell cycle- related proteins were determined with Western blotting. Results The expression of miR- 128 a were significantly up- regulated in HCC tissues as compared to the adjacent tissues（P〈0.05）. In cultured HCC cells, miR- 128 a promoted the cell proliferation and resulted in down- regulated RND3 mRNA and protein expressions by targeting RND3＇3＇UTR（P0.05） and also in the down-regulation of cyclin B1, cyclin D1 and CDK4 protein expressions. Conclusion miR-128 a is up-regulated in HCC and promotes HCC cell proliferation by targeting RND3.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2014年第10期1408-1413,共6页
Journal of Southern Medical University
基金
国家自然科学基金(81172056
81172586)~~
