Detection of dibenzo [a,l] pyrene-induced H-ras codon 61 mutant genes in preneoplastic SENCAR mouse skin using a new PCR-RFLP method

Dhrubajyoti Chakravarti, Paula Mailander, Jennifer Franzen, Sheila Higginbotham, Ercole L. Cavalieri, Eleanor G. Rogan

Research output: Contribution to journalArticle

50 Scopus citations

Abstract

One of the key events in tumor initiation in mouse skin is mutational activation of the H-ras gene. Papillomas induced by the most carcinogenic environmental polycyclic aromatic hydrocarbon (PAH), dibenzo[a,l]pyrene (DB[a,l]P), in SENCAR mouse skin contain a specific H-ras as codon 61 (CAA→CTA) mutation. We describe here detection of these mutations in preneoplastic skin by measuring the frequency of an induced XbaI RFLP, created by the mutation. Development of the PCR-XbaI RFLP method, sensitive enough to detect 1 codon 61 mutant allele among 10000 wild-type genes, is described. The results indicate that codon 61 mutations are induced 1 day (0.1%) after DB[a,l]P treatment on mouse skin, reach a high value (5%) by day 3, rapidly decline between days 7-9 and increase again during the clonal expansion of pre-papillomas into tumors. The detection of codon 61 mutations 1 day after DB[a,l]P exposure suggests that mutations occurred by pre-replication misrepair.

Original languageEnglish (US)
Pages (from-to)3203-3210
Number of pages8
JournalOncogene
Volume16
Issue number24
DOIs
StatePublished - Jun 18 1998

Keywords

  • DNA repair
  • Dibenzo[a,l]pyrene
  • Gleaning efficiency
  • In vivo mutation
  • Quantitative PCR
  • SENCAR mouse

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

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