Relationship between chemically induced Ha-ras mutation and transformation of BALB/c 3T3 cells: evidence for chemical-specific activation and cell type-specific recruitment of oncogene in transformation

Mol Carcinog. 1990;3(4):202-9. doi: 10.1002/mc.2940030407.

Abstract

BALB/c 3T3 cells were exposed to 7,12-dimethylbenz[a]anthracene (DMBA) and resultant transformed foci were analyzed for the presence of A182----T mutation at codon 61 of Ha-ras (a mutation found in many DMBA-induced animal tumors). None of the 30 independently cloned transformed cell lines contained such a mutation. In order to see whether DMBA is able to induce this mutation in BALB/c 3T3 cells, we developed a method sensitive enough to detect this specific mutation at the frequency of 10(-6). Employing this assay, we found time- and dose-dependent induction by DMBA of Ha-ras A182----T mutation in BALB/c 3T3 cells; for example, 2 wk after exposure to 100 micrograms/mL DMBA, 1.4 in 1 X 10(4) cells contained this specific mutation. On the other hand, other agents that also induce BALB/c 3T3 cell transformation, such as 3-methylcholanthrene (MCA), 12-O-tetradecanoylphorbol-13-acetate (TPA), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), or ultraviolet light, did not induce the mutation at detectable frequency (less than 10(-6)). These results suggest that DMBA efficiently induces Ha-ras mutation in BALB/c 3T3 cells but that this mutation is not recruited in the process of cell transformation. A hypothesis of carcinogen-specific mutation of Ha-ras gene and its tissue (cell type)-specific recruitment in carcinogenesis is proposed.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 9,10-Dimethyl-1,2-benzanthracene / toxicity
  • Animals
  • Antigens, Neoplasm / toxicity
  • Antigens, Tumor-Associated, Carbohydrate*
  • Base Sequence
  • Carcinogens*
  • Codon
  • DNA / drug effects*
  • Gene Expression Regulation / drug effects*
  • Gene Frequency
  • Genes, ras*
  • Methylnitronitrosoguanidine / toxicity
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Mutation*
  • Transformation, Genetic*
  • Tumor Cells, Cultured

Substances

  • Antigens, Neoplasm
  • Antigens, Tumor-Associated, Carbohydrate
  • Carcinogens
  • Codon
  • mucinous carcinoma-associated antigen
  • Methylnitronitrosoguanidine
  • 9,10-Dimethyl-1,2-benzanthracene
  • DNA