OBJECTIVETo construct a short hairpin RNA (shRNA) vector of the hypoxia inducible factor-1alpha (HIF-1alpha), determine its inhibitory effect on the expression of the HIF-1alpha gene in PC-3M cells, and investigate its application prospects in the treatment of prostate cancer.

METHODSWe designed and synthesized the shRNA template sequence specific against HIF-lalpha, inserted it into the vector psilencer 2.1-U6 to generate the plasmid psilencer-HIF, transfected the recombinant plasmid into prostate cancer cell line PC-3M cells and detected the transfection efficiency by cotransfection with the pEGFP vector as well as the expression of HIF-1alpha by RT-PCR and Western blot.

RESULTSThe DNA sequencing analysis showed a complete consistency of the recombinant plasmid psilencer-HIF with the design. Twenty-four hours after the transfection, the rate of transfected plasmid was about (89.26 +/- 4.72)% and the vector-mediated shRNA induced RNA interference (RNAi), while 48 hours transfection reduced the HIF-1alpha mRNA and protein levels by 82.09% and 81.61% respectively (P < 0.01) in PC-3M cells.

CONCLUSIONThe shRNA vector was successfully constructed, which can effectively suppress the expression of HIF-1alpha in prostate cancer cells.