Apoptosis in human papillomavirus16 E7-, but not E6-immortalized human uroepithelial cells.

We compared the ability of E6-, versus E7-, immortalized human uroepithelial cells (HUC) to undergo apoptosis in response to gamma radiation. Two independent HPV16 E6-immortalized cell lines, alphaE6#1 and alphaE6#2, that showed low or undetectable p53 levels, failed to undergo apoptosis in response to 18 Gray (Gy) gamma radiation as determined by DNA fragmentation. In contrast, two independent HPV16 E7-immortalized cell lines, alphaE7#1 and alphaE7#2, both of which showed stabilized wildtype p53, underwent apoptosis in the same experiment. Interestingly, both alphaE7#1 and alphaE7#2 showed constitutively elevated BAX and lowered BCL-2 levels, compared to either alphaE6#1 or alphaE6#2. However, elevated BAX and reduced BCL-2 per se were insufficient to trigger apoptosis, as apoptosis occurred only after exposure to gamma radiation. These results support a model in which HPV16 E7-immortalized cells are primed to undergo apoptosis, given an appropriate trigger. This apoptotic response was not observed in alphaE6/E7#1 cells which, like alphaE6-HUCs, showed low p53 levels, nor in late passage alphaE7#1 with spontaneously mutated TP53. These results suggest that E7 immortalization primes HUC for apoptosis in response to gamma radiation, and that this enhanced apoptotic response is p53 dependent.