He Jing, Mu Mimi, Luo Yulan, Wang Helong, Ma Hua, Guo Shujun, Fang Qiang, Qian Zhongqing, Lu Hezuo, Song Chuanwang
Department of Immunology, Bengbu Medical College, and Anhui Key Laboratory of Infection and Immunity at Bengbu Medical College, Bengbu, Anhui 233030, P.R. China.
Department of Microbiology and Parasitology, Bengbu Medical College, and Anhui Key Laboratory of Infection and Immunity at Bengbu Medical College, Bengbu, Anhui 233030, P.R. China.
Oncol Lett. 2019 Mar;17(3):2931-2936. doi: 10.3892/ol.2019.9925. Epub 2019 Jan 14.
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 (). The 3'-UTR sequence of the 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 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 , and it was revealed that miR-20b may directly target the 3'-untranslated region of the gene. Downregulation of partially reversed the anti-proliferative effect of miR-20b on H22 cells. In conclusion, miR-20b may promote H22 cell proliferation by targeting , providing a rationale for further study investigating novel therapeutic strategies for liver cancer.
微小RNA(miRNAs/miRs)是一类小的非编码RNA分子,与肿瘤的发生和发展密切相关。miR-20b在肝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。在本研究中,使用细胞计数试剂盒-8法检测H22小鼠肝癌细胞的增殖。利用MiRanda软件预测miR-20b与磷酸酶和张力蛋白同源物(PTEN)3'-非翻译区(3'-UTR)的结合位点。在H22细胞中通过聚合酶链反应扩增PTEN基因的3'-UTR序列。将重组质粒或空质粒与miR-20b模拟物或miR-20b乱序序列共转染入HeLa细胞,转染24小时后用双荧光素酶报告基因检测系统评估荧光素酶活性。在本研究中,miR-20b敲低显著抑制了H22小鼠肝癌细胞的增殖。此外,抑制miR-20b上调了PTEN的表达,并且发现miR-20b可能直接靶向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促进H22细胞增殖,为进一步研究肝癌的新型治疗策略提供了理论依据。