The present study was performed to gain insight into the role of p53 and p21(WAF1) on the cytotoxicity of the
purine analogue
cladribine (2-CdA) on
cancer cells.
Drug sensitivity, cell cycle distribution and
drug-induced cell death were compared in three lines derived from the
colorectal carcinoma HCT116: the p53+/+ cell line containing wild-type p53 and the p53-/- and p21(WAF1)-/- lines, in which both alleles of p53 or p21(WAF1) were deleted by homologous recombination, respectively. p53-/- and p21(WAF1)-/- cells were significantly more resistant to the cytotoxic effects of 2-CdA than the p53+/+ cells. p53+/+ cells and p21(WAF1)-/-, but not p53-/- cells, displayed wt-p53
protein accumulation and arrested in S-phase after exposure to 2-CdA.
mRNA analysis of the transporter hENT1 and of
enzymes involved in
drug metabolism did not show alterations which might explain a
drug-resistant phenotype in the p53-/- or p21(WAF1)-/- cells. Exposure of p53+/+ cells to 2-CdA resulted in expression of p21(WAF1)
mRNA and
protein, enhanced expression of uncleaved PARP-1, and a higher degree both of apoptosis and
necrosis than in p53-/- and p21(WAF1)-/- cells exposed to 2-CdA. Addition of the specific PARP-1 inhibitor 3-AB to 2-CdA-treated cells rendered p53+/+ cells resistant to this
drug. Bax levels were reduced in the p53-/- while they increased in the p53+/+ line and remained stable in the p21(WAF1)-/- cells. We conclude that p53 and p21(WAF1) status of
cancer cells influences their sensitivity to 2-CdA cytotoxicity. This may involve alterations in the apoptotic cascade as well as in PARP-1-dependent cell death.