Over‐expression of hypoxia‐inducible factor‐1α increases the invasive potency of LNCaP cells in vitro

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Abstract
OBJECTIVE: To evaluate the effect of hypoxia‐inducible factor‐1α (HIF‐1α) over‐expression on the invasion‐associated proteins in human prostate cancer cells, as HIF‐1α 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.MATERIALS AND METHODS: Prostate cancer (LNCaP) cells were transfected with recombinant plasmid pcDNA3.1(–)/HIF‐1α and pcDNA3.1(–) control vector using a commercial system, designated as LNCaP/HIF‐1α 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‐1α in LNCaP/HIF‐1α was distinctly higher than that in LNCaP/pcDNA3.1 and LNCaP. Many more cells of LNCaP/HIF‐1α 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‐1α, whereas the expression of E‐cadherin was down‐regulated.CONCLUSION: These results show that over‐expression of HIF‐1α 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‐1α in human prostate cancer cells.