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 Cavalieri, Eleanor G Rogan

Research output: Contribution to journalArticle

50 Citations (Scopus)

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

Fingerprint

Inbred SENCAR Mouse
Codon
Restriction Fragment Length Polymorphisms
Polymerase Chain Reaction
Skin
Mutation
Genes
Papilloma
ras Genes
Polycyclic Aromatic Hydrocarbons
dibenzo(a,l)pyrene
Neoplasms
Alleles

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

Cite this

Detection of dibenzo [a,l] pyrene-induced H-ras codon 61 mutant genes in preneoplastic SENCAR mouse skin using a new PCR-RFLP method. / Chakravarti, Dhrubajyoti; Mailander, Paula; Franzen, Jennifer; Higginbotham, Sheila; Cavalieri, Ercole; Rogan, Eleanor G.

In: Oncogene, Vol. 16, No. 24, 18.06.1998, p. 3203-3210.

Research output: Contribution to journalArticle

Chakravarti, Dhrubajyoti ; Mailander, Paula ; Franzen, Jennifer ; Higginbotham, Sheila ; Cavalieri, Ercole ; Rogan, Eleanor G. / Detection of dibenzo [a,l] pyrene-induced H-ras codon 61 mutant genes in preneoplastic SENCAR mouse skin using a new PCR-RFLP method. In: Oncogene. 1998 ; Vol. 16, No. 24. pp. 3203-3210.
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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.",
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