Over-expression of Hypoxia-inducible factor 1 alpha increases invasive potency of LNCaP cells in vitro

Objective To evaluate the effect of hypoxia-inducible factor-la (HIF1α) over-expression on the invasive potency of human prostate cancer cell. Methods Human prostate cancer cell of the line LNCaP were cultured and transfected by the recombinant plasmid pcDNA3. 1 (-)-HIF-1αcontaining the gene HIF1αwith Lipofectamine 2000 system. The positive clone cells were selected by C418 and confirmed by Western blotting and immunofluorescence staining (LNCaP/ HIF1αcells). Transwell chambers with polycarbonate filter were coated by 100μl Matrigel at 1:20 dilution in serum-free medium. LNCaP cell suspension and LNCaP/ HIF1αcell suspension were inoculated into the Transwell chambers respectively for 24 hours to analyze the invasive potency. Western blotting was used to detect the expression of E-cadherin, vimentin, matrix metalloproteinase-2 ( MMP-2), cathepsin D, and urokinase-type plasminogen activator receptor (uPAR). Results The expression level of HIF1αin the LNCaP/HIF1αcells was distinctly higher than that in the LNCaP cells. The numbers of LNCaP-HIF1αcells penetrating through the Transwell polycarbonate filter was 4.6±0.4×104 , significantly higher than that of the LNCaP cells (3.2±0.3×104, P 0.01). The expressions of vimentin, MMP-2, cathepsin D, and uPAR were all up-regulated in LNCaP-HIF1αcells than those of the LNCaP cells. Whereas, the expression of E-cadherin was down-regulated in the LNCaP-HIF1αcells. Conclusion Over-expression of HIF-1αstimulates the invasion potency of human prostate carcinoma cell. The expression of E-cadherin, vimentin, MMP-2, cathepsin D, and uPAR, all playing an established role in the invasion of tumor, can be regulated by HIF-1 in human prostate cancer cell.