Influence of p53 and p21(WAF1) expression on sensitivity of cancer cells to cladribine

Biochem Pharmacol. 2003 Jan 1;65(1):121-9. doi: 10.1016/s0006-2952(02)01448-x.


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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Apoptosis
  • Blotting, Northern
  • Blotting, Western
  • Cell Cycle / drug effects
  • Cell Survival / drug effects
  • Cladribine / pharmacology*
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins / metabolism*
  • Drug Screening Assays, Antitumor
  • Gene Expression / drug effects
  • Humans
  • Necrosis
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tumor Cells, Cultured
  • Tumor Suppressor Protein p53 / metabolism*


  • Antineoplastic Agents
  • CDKN1A protein, human
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins
  • Tumor Suppressor Protein p53
  • Cladribine