MicroRNA-20b promotes proliferation of H22 hepatocellular carcinoma cells by targeting PTEN
MicroRNA-20b promotes proliferation of H22 hepatocellular carcinoma cells by targeting PTEN
复制标题
DOI：
10.3892/ol.2019.9925
复制
发表时间：
2019-03-01
期刊：
影响因子：
2.9
通讯作者：
Song, Chuanwang
中科院分区：
文献类型：
--
作者：
He, Jing;Mu, Mimi;Song, Chuanwang
MicroRNAs(miRNAs/miRs)是一种小的、非编码的RNA分子，与肿瘤的发生发展密切相关。MIR-20b在肝细胞癌细胞系和组织中过表达。然而，miR-20b是否能促进肝癌细胞的增殖尚不清楚。本研究采用细胞计数试剂盒-8法检测H22小鼠肝癌细胞的增殖情况。用Miranda软件预测miR-20b与磷酸酶张力蛋白同源物(PTEN)3‘-非翻译区(3’-UTR)的结合位点。用聚合酶链式反应在H22细胞中扩增出PTEN基因的3‘端非编码区序列。将重组质粒或空质粒与miR-20b模拟物或miR-20b置乱共转染HeLa细胞，24 h后用双荧光素酶((R)Reporter Assay System)检测荧光素酶活性。在本研究中，miR-20b基因敲除显著抑制了H22小鼠肝癌细胞的增殖。此外，抑制miR-20b还上调了PTEN的表达，提示miR-20b可能直接针对PTEN基因的3‘非翻译区。PTEN的下调部分逆转了miR-20b对H22细胞的抗增殖作用。总之，miR-20b可能通过靶向PTEN来促进H22细胞的增殖，为进一步研究肝癌的新治疗策略提供了理论基础。
MicroRNAs (miRNAs/miRs) are small, noncoding RNA molecules that are closely associated with the occurrence and development of tumors. miR-20b is overexpressed in hepatocellular carcinoma cell lines and tissues. However, it is not clear whether miR-20b can promote the proliferation of hepatocellular carcinoma cells. In the present study, the proliferation of H22 mouse hepatocellular carcinoma cells was detected using the Cell Counting Kit-8 assay. MiRanda software was used to predict the binding sites of miR-20b to the 3 '-untranslated region (3 '-UTR) of phosphatase and tensin homolog (PTEN). The 3 '-UTR sequence of the PTEN gene was amplified using the polymerase chain reaction in H22 cells. The recombinant plasmid or empty plasmid was co-transfected with miR-20b mimics or miR-20b scramble into HeLa cells, and luciferase activity was assessed by Dual-Luciferase((R)) Reporter Assay System 24 h post-transfection. In the present study, miR-20b knockdown significantly inhibited the proliferation of H22 mouse hepatocellular carcinoma cells. In addition, miR-20b inhibition upregulated the expression of PTEN, and it was revealed that miR-20b may directly target the 3 '-untranslated region of the PTEN gene. Downregulation of PTEN partially reversed the anti-proliferative effect of miR-20b on H22 cells. In conclusion, miR-20b may promote H22 cell proliferation by targeting PTEN, providing a rationale for further study investigating novel therapeutic strategies for liver cancer.