title: [Inhibitory effect of miRNA silencing hypoxia-inducible factor alpha subunit gene on the proliferation of HepG2 cells].
abstract: OBJECTIVE
  To investigate the effect of miRNA silencing HIF-1α gene on the proliferation of HepG2 cells.
  
  
  METHODS
  The eukaryotic expression plasmids of HIF-1α miRNA and report gene containing hypoxia-reponse element were constructed and transfected into HepG2 cells. The expressions of HIF-1α gene and protein were determined by real time-PCR and Western blotting. The expressions of HIF-1α, vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) were quantitatively detected by ELISA. The alterations of cell cycles and apoptosis rate were quantitatively measured by flow cytometry and Annexin V-FITC/PI double dyeing assay.
  
  
  RESULTS
  72 h after transfection the down regulations of HIF-1α mRNA and protein were 87% and 56% respectively, and the decrease of target gene was 46% in the report gene, 54% in VEGF and 36% in Ang-2, respectively. The apoptotic ratio of HepG2 cells was 22.46+/-0.61% (P < 0.01). The cell cycle changed greatly at the ratio of G1 (61.49+/-1.12%) and S (22.40+/-0.58%, P < 0.01). After being combined with doxorubicin, the apoptotic ratio increased to 36.99+/-0.88% and the ratios of G1 and S phases were upregulated to 65.68+/-0.91% and 19.47+/-1.34% respectively.
  
  
  CONCLUSIONS
  HIF-1α miRNA or / and doxorubicin can regulate the growth cycles of HepG2 cells, promote the cell apoptosis and inhibit the cell proliferation.
authors.affiliation: Department of Diagnostics, Medical School of Nantong University, Nantong Jiangsu Province 226001, China.
authors.forename: Zhi-zhen
authors.initials: ZZ
authors.lastname: Dong
authors.forename: Deng-fu
authors.initials: DF
authors.lastname: Yao
authors.forename: Shan-shan
authors.initials: SS
authors.lastname: Li
authors.forename: Min
authors.initials: M
authors.lastname: Yao
authors.forename: Dan-dan
authors.initials: DD
authors.lastname: Yu
authors.forename: Ning-hua
authors.initials: NH
authors.lastname: Yao
authors.forename: Ya-jie
authors.initials: YJ
authors.lastname: Qian
authors.forename: Li-wei
authors.initials: LW
authors.lastname: Qiu
mesh_terms: D017209:Apoptosis; D002453:Cell Cycle; D049109:Cell Proliferation; D020868:Gene Silencing; D056945:Hep G2 Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D014162:Transfection
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D035683:MicroRNAs; D012333:RNA, Messenger