MicroRNA-548a-5p promotes proliferation and inhibits apoptosis in hepatocellular carcinoma cells by targeting Tg737

• Published in: World journal of gastroenterology : WJG Vol. 22; no. 23; pp. 5364 - 5373
• Main Authors: Zhao, Ge, Wang, Ting, Huang, Qi-Ke, Pu, Meng, Sun, Wei, Zhang, Zhuo-Chao, Ling, Rui, Tao, Kai-Shan
• Format: Journal Article
• Language: English
• Published: United States Baishideng Publishing Group Inc 21.06.2016
• Subjects: 3' Untranslated Regions; Animals; Apoptosis - genetics; Basic Study; Carcinogenesis - genetics; Carcinogenesis - metabolism; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - metabolism; Cell Count; Cell Cycle; Cell Proliferation - genetics; Colony-Forming Units Assay; Down-Regulation; Gene Expression Regulation, Neoplastic; Hep G2 Cells; Humans; Liver Neoplasms - genetics; Liver Neoplasms - metabolism; Mice; Mice, Inbred BALB C; Mice, Nude; MicroRNAs - metabolism; Real-Time Polymerase Chain Reaction; Tumor Suppressor Proteins - metabolism; Up-Regulation; Xenograft Model Antitumor Assays; Hepatocellular carcinoma cells; microRNA-548a-5p; Proliferation; Tg737; Apoptosis
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.v22.i23.5364

To investigate whether Tg737 is regulated by microRNA-548a-5p (miR-548a-5p), and correlates with hepatocellular carcinoma (HCC) cell proliferation and apoptosis. Assays of loss of function of Tg737 were performed by the colony formation assay, CCK assay and cell cycle assay in HCC cell lines. The interaction between miR-548a-5p and its downstream target, Tg737, was evaluated by a dual-luciferase reporter assay and quantitative real-time polymerase chain reaction. Tg737 was then up-regulated in HCC cells to evaluate its effect on miR-548a-5p regulation. HepG2 cells stably overexpressing miR-548a-5p or miR-control were also subcutaneously inoculated into nude mice to evaluate the effect of miR-548a-5p up-regulation on in vivo tumor growth. As the final step, the effect of miR-548a-5p on the apoptosis induced by cisplatin was evaluated by flow cytometry. Down-regulation of Tg737, which is a target gene of miR-548a-5p, accelerated HCC cell proliferation, and miR-548a-5p promoted HCC cell proliferation in vitro and in vivo. Like the down-regulation of Tg737, overexpression of miR-548a-5p in HCC cell lines promoted cell proliferation, increased colony forming ability and hampered cell apoptosis. In addition, miR-548a-5p overexpression increased HCC cell growth in vivo. MiR-548a-5p down-regulated Tg737 expression through direct contact with its 3' untranslated region (UTR), and miR-548a-5p expression was negatively correlated with Tg737 levels in HCC specimens. Restoring Tg737 (without the 3'UTR) significantly hampered miR-548a-5p induced cell proliferation, and rescued the miR-548a-5p induced cell proliferation inhibition and apoptosis induced by cisplatin. MiR-548a-5p negatively regulates the tumor inhibitor gene Tg737 and promotes tumorigenesis in vitro and in vivo, indicating its potential as a novel therapeutic target for HCC.