Role of the mismatch repair system and p53 in the clastogenicity and cytotoxicity induced by bleomycin

Patrizia Vernole, Bruna Tedeschi, Lucio Tentori, Lauretta Levati, Gabriella Argentin, Rosadele Cicchetti, Olindo Forini, Grazia Graziani, Stefania D'Atri

Research output: Contribution to journalArticle

Abstract

The mismatch repair (MMR) system and p53 protein play a pivotal role in maintaining genomic stability and modulate cell chemosensitivity. Aim of this study was to examine the effects of either MMR-deficiency or p53 inactivation, or both, on cellular responses to bleomycin. The MMR-deficient colon carcinoma cell line HCT116 and its MMR-proficient subline HCT116/3-6, both expressing wild-type p53, were transfected with an expression vector encoding a dominant-negative p53 mutant, or with the empty vector. Four transfected clones, having the following phenotypes, MMR-proficient/p53 wild-type, MMR-proficient/p53 mutant, MMR-deficient/p53 wild-type, MMR-deficient/p53 mutant, were subjected to treatment with bleomycin. Loss of MMR function alone was associated with increased resistance to apoptosis, chromosomal damage and inhibition of colony formation caused by bleomycin. Loss of p53 alone resulted in abrogation of G1 arrest and increased sensitivity to apoptosis and chromosomal damage induced by the drug, but did not affect clonogenic survival after bleomycin treatment. Disabling both p53 and MMR function led to abrogation of G1 arrest and to a moderate impairment of drug-induced apoptosis. Chromosomal damage was reduced in the MMR-deficient/p53 mutant clone with respect to the MMR-proficient/p53 wild-type one, when evaluated 48 h after bleomycin treatment, but was comparable in both clones 96 h after drug exposure. Clonogenic survival of the MMR-deficient/p53 mutant clone was similar to that of the MMR-deficient/p53 wild-type one. The effects of MMR-deficiency on cellular responses to bleomycin were confirmed using the MMR-proficient lymphoblastoid cell line TK6 and its MMR-deficient subline MT1, both expressing wild-type p53. In conclusion, our data show that loss of MMR and p53 function exerts opposite and independent effects on apoptosis and chromosomal damage induced by bleomycin. Moreover, inactivation of MMR confers resistance to the cytotoxic activity of the anticancer agent in cells expressing either wild-type or mutant p53.

Original languageEnglish
Pages (from-to)63-77
Number of pages15
JournalMutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
Volume594
Issue number1-2
DOIs
Publication statusPublished - Feb 22 2006

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DNA Mismatch Repair
Bleomycin
Clone Cells
Apoptosis
Pharmaceutical Preparations
Cell Line
Genomic Instability

Keywords

  • Apoptosis
  • Bleomycin
  • Cell cycle
  • Chromosomal damage
  • Mismatch repair
  • p53

ASJC Scopus subject areas

  • Molecular Biology
  • Health, Toxicology and Mutagenesis

Cite this

Role of the mismatch repair system and p53 in the clastogenicity and cytotoxicity induced by bleomycin. / Vernole, Patrizia; Tedeschi, Bruna; Tentori, Lucio; Levati, Lauretta; Argentin, Gabriella; Cicchetti, Rosadele; Forini, Olindo; Graziani, Grazia; D'Atri, Stefania.

In: Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, Vol. 594, No. 1-2, 22.02.2006, p. 63-77.

Research output: Contribution to journalArticle

Vernole, Patrizia ; Tedeschi, Bruna ; Tentori, Lucio ; Levati, Lauretta ; Argentin, Gabriella ; Cicchetti, Rosadele ; Forini, Olindo ; Graziani, Grazia ; D'Atri, Stefania. / Role of the mismatch repair system and p53 in the clastogenicity and cytotoxicity induced by bleomycin. In: Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis. 2006 ; Vol. 594, No. 1-2. pp. 63-77.
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AU - Argentin, Gabriella

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