Inhibition of JNK-1 by small interfering RNA induces apoptotic signaling in PC-3 prostate cancer cells.
ABSTRACT Previous studies have shown that c-Jun-N-terminal kinase-1 (JNK-1) is involved in the transformation of primary fibroblasts and plays a role in tumor cell growth. A number of observations suggest that JNK-1 is a growth promoting factor in prostate cancer cells and blocking its function may induce apoptosis. To test this further, we used a small interfering RNA (siRNA) against JNK-1 mRNA that efficiently inhibits JNK-1 expression in the prostate cancer cell line, PC-3. The application of siRNA against JNK-1 decreased the expression of JNK-1 and affected the expression of p21, XIAP and Bcl-2, but had no effect on the expression of VEGF. In contrast, a control scramble siRNA did not affect the expression of the above indicated proteins. The downregulation of JNK-1 expression at both the mRNA and protein levels was detected by reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis. Cell proliferation inhibition rates were determined by the MTT assay. The effect of JNK-1-siRNA on cell cycle distribution and cell apoptosis was determined by flow cytometry, DNA fragmentation and caspase activity. Our data showed that siRNA against JNK-1 mRNA, could efficiently suppress the expression of JNK-1 in PC-3 cells. After 5 days of transfection, the cell death rate was 52%, the apoptotic rate 26% and the viability rate 22%. In conclusion, downregulation of JNK-1 expression by siRNA against JNK-1 mRNA induces apoptotic signaling in prostate cancer PC-3 cells. The use of siRNA against JNK-1 as a novel approach to cancer therapy deserves further investigation.