Lentivirus-Mediated Overexpression of MicroRNA-199a Inhibits Cell Proliferation of Human Hepatocellular Carcinoma

• Published in: Cell biochemistry and biophysics 2012, Vol.62 (1), p.237-244
• Main Authors: Jia, Xiao Qin, Cheng, Hai Qing, Qian, Xu, Bian, Chuan Xiu, Shi, Zhu Mei, Zhang, Jian Ping, Jiang, Bing Hua, Feng, Zhen Qing
• Format: Article
• Language: English
• Subjects: 3' Untranslated Regions; Animals; Biochemistry; Biological and Medical Physics; Biomedical and Life Sciences; Biophysics; Biotechnology; Carcinoma, Hepatocellular - pathology; Cell Biology; Cell Hypoxia; Cell Line, Tumor; Cell Proliferation; Expression vectors; Gene therapy; Genetic Vectors - genetics; Hepatocellular carcinoma; Hepatocytes; Humans; Hypoxia; Hypoxia-inducible factor 1 alpha; Hypoxia-Inducible Factor 1, alpha Subunit - antagonists & inhibitors; Hypoxia-Inducible Factor 1, alpha Subunit - metabolism; Lentivirus - genetics; Life Sciences; Liver Neoplasms - pathology; Mice; Mice, Nude; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; Pharmacology/Toxicology; Polymerase chain reaction; Translational Biomedical Research; Transplantation, Heterologous; Tumorigenesis
• ISSN: 1085-9195; 1559-0283
• DOI: 10.1007/s12013-011-9263-8

microRNA-199a (miR-199a) is a highly conserved miRNA, always deregulated in numerous human tumors. The results of microarray analysis indicated that miR-199a was frequently downregulated in hepatocellular carcinoma (HCC). The expression levels of miR-199a in 11 pairs of matched HCC neoplastic and adjacent non-neoplastic tissues, 5 HCC cell lines and liver cell line L02 were examined by quantitative real-time PCR analysis. We found miR-199a was significantly down-regulated in HCC tissues when compared with pair-matched adjacent non-tumor tissues. We also found the expression level of miR-199a was also substantially decreased in several human HCC cell lines including SMMC-7721, BEL-7402, BEL-7701, MHCC97H, and HepG2. To investigate the role of miR-199a in tumorigenesis, we developed a lentiviral vector for the expression of pre-miR-199a (Lenti-miR-199a). Lenti-miR-199a inhibited HCC cell proliferation in vitro and in vivo. Compared to parental cells or cells transfected with a control vector, the overexpression of microRNA-199a in the HCC cell lines HepG2 stably was showed to reduce cell proliferation in vitro and in vivo. Luciferase reporter assay revealed the regulation of miR-199a on 3’-UTR of HIF-1α. Further investigation confirmed that miR-199a significantly reduced the endogenous protein level of HIF-1α in hypoxia. MiR-199a inhibits cell proliferation in vitro and in vivo partly through down-regulation of HIF-1α in human HCC. Thus, these studies provide an important new insight into the pathogenesis of human HCC and it may open a new perspective for the development of effective gene therapy for human HCC.