Emergence of p53 mutant cisplatin-resistant ovarian carcinoma cells following drug exposure: Spontaneously mutant selection

Sabina C. Righetti, Paola Perego, Elisabetta Corna, Marco A. Pierotti, Franco Zunino

Research output: Contribution to journalArticle

Abstract

We have previously shown that p53 mutations are associated with cisplatin resistance in ovarian carcinoma IGROV-1/Pt 1 cells. The relationship between p53 status and the development of resistance has not been completely elucidated; in particular, the biological mechanisms behind the acquired drug-resistant p53-mutant phenotype were not clearly explained. Thus, in this study, we investigated whether the p53 mutations found in IGROV-1/Pt 1 cells (270 and 282 codons) resulted from selection, under the selective pressure of the cytotoxic treatment, of a spontaneously mutant cell population preexistent in the cisplatin-sensitive parental cell line (IGROV- 1) or were induced by drug (genotoxic) treatment. For this purpose, an allele-specific PCR approach was used. Primers carrying the desired mutations (T→A codon 270, C→T codon 282) in the 3' terminus, and the corresponding wild-type primers were used to amplify genomic DNA from the original IGROV-1 cell line used to select the mutant IGROV-1/Pt 1. To increase sensitivity, we hybridized blots of the PCRs with the radiolabeled PCR fragment from IGROV- 1/Pt 1. Amplification was obtained for IGROV-1 DNA with the mutated allele- specific primers, indicating the preexistence of a mutated population in the IGROV-1 cell line. Titration experiments suggested that the frequency of the mutated alleles was

Original languageEnglish
Pages (from-to)473-478
Number of pages6
JournalCell Growth and Differentiation
Volume10
Issue number7
Publication statusPublished - Jul 1999

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

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