Yang Xiaoyan, Yu Jia, Yin Jie, Xiang Qiong, Tang Huifang, Lei Xiaoyong
Institute of Pharmacy and Pharmacology, The First Affilitated Hospital, University of South China Hengyang, P.R. China.
Pharmazie. 2012 Jul;67(7):645-51.
Recently, we have reported tissue- and stage-specific expression of miR-195 in human hepatocellular carcinoma cells and so far, not many reports discuss the function of this microRNA (miRNA). Expression profiling of miRNAs revealed a limited set of miRNAs with altered expression in drug resistant hepatocellular carcinoma cell line BEL-7402/5-FU compared to its parental BEL-7402 cell line. Real-time PCR confirmed down-regulation of miR-195 in BEL-7402/5-FU cells. Western blots were performed to determine protein levels of LATS2, P53 and CDK2. MTT analysed the cell proliferation activity. Flow cytometry were performed to determine apoptosis rate. Up-regulation of miR-195 increased expression of LATS2 and increased apoptosis of HCC cells, while Anti-miR-195 treatment inhibited expression of LATS2. miR-195 over-expression inhibited the luciferase activity of a LATS2 3' untranslated region-based reporter construct in BEL-7402/5-FU cells. These results indicate that miR-195 could increase cell apoptosis by targeting LATS2 in hepatocellular carcinoma cells.
最近,我们报道了miR-195在人肝癌细胞中的组织和阶段特异性表达,到目前为止,关于这种微小RNA(miRNA)功能的报道并不多。miRNA表达谱分析显示,与亲本BEL-7402细胞系相比,耐药肝癌细胞系BEL-7402/5-FU中表达改变的miRNA数量有限。实时PCR证实BEL-7402/5-FU细胞中miR-195表达下调。进行蛋白质印迹以确定LATS2、P53和CDK2的蛋白质水平。MTT分析细胞增殖活性。进行流式细胞术以确定凋亡率。miR-195的上调增加了LATS2的表达并增加了肝癌细胞的凋亡,而抗miR-195处理则抑制了LATS2的表达。miR-195的过表达抑制了基于BEL-7402/5-FU细胞中LATS2 3'非翻译区的报告基因构建体的荧光素酶活性。这些结果表明,miR-195可通过靶向肝癌细胞中的LATS2来增加细胞凋亡。