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Related Experiment Videos

Consistent oncogene methylation changes in epithelial cells chemically transformed in vitro.

M J Mass1, N S Schorschinsky, J A Lasley

  • 1Genetic Toxicology Division, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711.

Biochemical and Biophysical Research Communications
|October 31, 1989
PubMed
Summary

DNA methylation patterns in oncogenes c-myc and Ha-ras are altered in rat tracheal cells exposed to carcinogens. These changes, observed via restriction digest patterns, suggest common mechanisms for polycyclic hydrocarbons and tumor promoters.

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Area of Science:

  • Molecular biology
  • Oncology
  • Cellular toxicology

Background:

  • Investigating oncogene alterations in epithelial cells is crucial for understanding carcinogenesis.
  • Polycyclic aromatic hydrocarbons and tumor promoters are known environmental carcinogens.
  • Specific DNA methylation changes in oncogenes like c-myc and Ha-ras are linked to cancer development.

Purpose of the Study:

  • To analyze restriction digest patterns of CCGG sequences in Ki-ras, Ha-ras, and c-myc oncogenes.
  • To compare these patterns in normal rat tracheal epithelial cells versus cells transformed by chemical carcinogens.
  • To identify molecular alterations indicative of oncogene activation by specific agents.

Main Methods:

  • Utilized restriction enzyme digestion (Hpa II) to examine CCGG sequences.

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  • Employed Southern blot hybridization to detect alterations in oncogene restriction patterns.
  • Compared molecular weight and presence/absence of hybridizing bands between normal and transformed cells.

    • Main Results:

    • Altered Hpa II restriction patterns were observed in c-myc and Ha-ras oncogenes in transformed cell lines.
    • These alterations included changes in hybridizing band molecular weights and band loss.
    • Ki-ras oncogene exhibited hypermethylation across all cell types, including normal controls.

    Conclusions:

    • In vitro transformation of rat tracheal epithelial cells by polycyclic hydrocarbons and TPA induces specific alterations in c-myc and Ha-ras oncogene methylation.
    • The observed molecular changes in oncogenes have not been previously reported for these cell transformation models.
    • Findings suggest shared molecular mechanisms of action for diverse carcinogenic agents targeting epithelial cells.