title: Over-expression of hypoxia-inducible factor-1alpha increases the invasive potency of LNCaP cells in vitro.
abstract: OBJECTIVE
  To evaluate the effect of hypoxia-inducible factor-1alpha (HIF-1alpha) over-expression on the invasion-associated proteins in human prostate cancer cells, as HIF-1alpha is a transcriptional factor that could activate genes involved in the response to hypoxia, but might also enhance the invasive potency of prostate cancer cells.
  
  
  METHODS
  Prostate cancer (LNCaP) cells were transfected with recombinant plasmid pcDNA3.1(-)/HIF-1alpha and pcDNA3.1(-) control vector using a commercial system, designated as LNCaP/HIF-1alpha and LNCaP/pcDNA3.1, respectively. The positive clones were selected with G418 and confirmed by Western blot and indirect immunofluorescence labelling. A polycarbonate filter, coated with a matrix gel, was used to analyse the invasive potency. The expression of E-cadherin, vimentin, cathepsin D, matrix metalloproteinase (MMP2) and urokinase plasminogen activator receptor (uPAR) was detected by Western blot.
  
  
  RESULTS
  The expression level of HIF-1alpha in LNCaP/HIF-1alpha was distinctly higher than that in LNCaP/pcDNA3.1 and LNCaP. Many more cells of LNCaP/HIF-1alpha penetrated through the polycarbonate filter than those of LNCaP/pcDNA3.1 and LNCaP. Compared with the LNCaP/pcDNA3.1 and LNCaP cells, the expression of vimentin, cathepsin D, MMP-2 and uPAR were up-regulated in LNCaP/HIF-1alpha, whereas the expression of E-cadherin was down-regulated.
  
  
  CONCLUSIONS
  These results show that over-expression of HIF-1alpha directly stimulates the invasive potency of human prostate carcinoma LNCaP cells through the matrix gel. The expression of E-cadherin, vimentin, cathepsin D, MMP-2 and uPAR, which are proteins with established roles in the pathophysiology of invasion, could be regulated by HIF-1alpha in human prostate cancer cells.
authors.affiliation: Department of Urology, First Hospital of Xi'an Jiaotong University, Xi'an Jiaotong University, Ministry of Education, Xi'an, People's Republic of China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Luo
authors.forename: Da-Lin
authors.initials: DL
authors.lastname: He
authors.forename: Liang
authors.initials: L
authors.lastname: Ning
authors.forename: Shu-Lin
authors.initials: SL
authors.lastname: Shen
authors.forename: Lei
authors.initials: L
authors.lastname: Li
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.forename: Haiyen E
authors.initials: HE
authors.lastname: Zhau
authors.forename: Leland W K
authors.initials: LW
authors.lastname: Chung
mesh_terms: D015153:Blotting, Western; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism*; D008297:Male; D009361:Neoplasm Invasiveness / Q000473:pathology*; D011471:Prostatic Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit