| Abstract: | In this study we evaluated the effect of over-expression of the bcl-2 gene, a potent apoptosis suppressor, on radiation-induced apoptotic cell death in 2 human prostate cancer cell lines, androgen-independent PC-3 cells and androgen-sensitive LNCaP cells. Cells were transfected with the bcl-2 gene and bcl-2 transfectant clones isolated under neomycin selection; bcl-2 gene integration and level of mRNA and protein expression in the cloned transfectants were examined by Southern, Northern and Western blot analyses, respectively. Parental, neo control and bcl-2-expressing cells were exposed to single or fractionated doses of ionizing irradiation, and the cellular response to radiation was determined at 24, 48 and 72 hr post-irradiation, on the basis of: (i) loss of cell viability, (ii) clonogenic survival and (iii) induction of apoptotic DNA fragmentation. At 24 hr post-irradiation all cell lines, i.e., parental and bcl-2 transfectants, failed to form colonies, though the majority of bcl-2-expressing cells did not exhibit apoptotic morphology; bcl-2 over-expression in both cell lines reduced apoptosis 48 hr post-irradiation from 20–25% to 5% at a dose of 2,000 cGy. By 72 hr, bcl-2 over-expression afforded a 3-fold protection from radiation-induced apoptosis. There was no significant difference, however, in the clonogenic survival of the parental and bcl-2-expressing cells. Furthermore, there was a 24 hr delay in induction of the apoptosis marker gene SGP-2/TRPM-2 in the bcl-2-expressing cells, co-incidental with the delay in apoptotic DNA fragmentation. Int. J. Cancer, 70:341–348, 1997. © 1997 Wiley-Liss, Inc. |
