Treatment of prostate cancer cells with adenoviral vector-mediated antisense RNA using androgen-dependent and androgen-independent promoters.

The present study was designed to develop a novel antisense approach to prostate cancer therapy. We constructed ODC/AdoMetDC double antisense RNA recombinant adenovirus mediated by a prostate-specific androgen-dependent promoter (pADxsi-P2-AdoMetDC-ODC-PolyA) or a prostate-specific androgen-independent promoter (pADxsi-P1-AdoMetDC-ODC-PolyA). Western blot analysis was performed to detect the ODC and AdoMetDC protein levels. The growth curves for each group of cells were determined by MTT assay. Flow cytometry was conducted to detect cell apoptosis, proliferation, and cell cycle distribution in order to demonstrate the effects of the recombinant adenoviruses on the prostate cancer cells. RT-PCR, Western blotting, MTT, and tumor growth inhibition assay in nude mice demonstrated that pADxsi-P1-AdoMetDC-ODC-PolyA and pADxsi-P2-AdoMetDC-ODC-PolyA exhibited inhibitory effects on cell proliferation at both the gene and protein levels. Meanwhile, inhibiting effects of the pADxsi-P2-AdoMetDC-ODC-PolyA is more profound than those of the pADxsi-P1-AdoMetDC-ODC-PolyA vector. Both of the adenoviral vectors exhibited significant inhibitory effects on the growth of prostate tumors, and the inhibitory effects of the pADxsi-P2-AdoMetDC-ODC-PolyA vector were stronger than those of the pADxsi-P1-AdoMetDC-ODC-PolyA vector.