title: Lentivirus-mediated overexpression of microRNA-199a inhibits cell proliferation of human hepatocellular carcinoma.
abstract: 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.
authors.affiliation: Cancer Center & Department of Pathology, Nanjing Medical University, Nanjing, China.
authors.forename: Xiao Qin
authors.initials: XQ
authors.lastname: Jia
authors.forename: Hai Qing
authors.initials: HQ
authors.lastname: Cheng
authors.forename: Xu
authors.initials: X
authors.lastname: Qian
authors.forename: Chuan Xiu
authors.initials: CX
authors.lastname: Bian
authors.forename: Zhu Mei
authors.initials: ZM
authors.lastname: Shi
authors.forename: Jian Ping
authors.initials: JP
authors.lastname: Zhang
authors.forename: Bing Hua
authors.initials: BH
authors.lastname: Jiang
authors.forename: Zhen Qing
authors.initials: ZQ
authors.lastname: Feng
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000037:antagonists & inhibitors / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D008113:Liver Neoplasms / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism*; D014183:Transplantation, Heterologous
chemical_list: D020413:3' Untranslated Regions; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D035683:MicroRNAs; C531408:mirn199 microRNA, human