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Antipain-induced suppression of oncogene expression in H-ras-transformed NIH3T3 cells

L R Cox1, J Motz, W Troll

  • 1Institute of Environmental Medicine, New York University Medical Center, New York 10016.

Cancer Research
|September 15, 1991
PubMed

Insights

Antipain (AP) suppresses H-ras oncogene-induced cell transformation by reducing expression of both transfected and endogenous oncogenes. This inhibition of oncogene expression is linked to the loss of the transformed cell phenotype.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Oncology

Background:

  • Cell transformation is a critical step in cancer development.
  • Oncogenes, such as H-ras, play a key role in driving cellular transformation.
  • Investigating compounds that can modulate oncogene expression is crucial for cancer research.

Purpose of the Study:

  • To investigate the effect of Antipain (AP) on H-ras oncogene-induced NIH3T3 cell transformation.
  • To determine if AP's inhibitory effect on transformation is mediated by alterations in oncogene expression.
  • To quantify changes in both exogenous H-ras and endogenous c-myc expression.

Main Methods:

  • NIH3T3 cells were co-transfected with an activated H-ras oncogene and a selectable marker gene (aph).
  • Cells were treated with Antipain (AP) at 50 micrograms/ml.
  • Gene expression was quantified using Northern blot analysis.
  • Cellular transformation was assessed by the ability to grow in soft agar.
  • Southern blot analysis was performed to assess gene incorporation and copy number.

Main Results:

  • Antipain (AP) inhibited the transformed phenotype in 50% of geneticin-resistant colonies.
  • AP treatment led to a 4-fold decrease in the expression of the transfected H-ras gene.
  • Expression of the endogenous c-myc oncogene decreased by approximately 2.5-fold in AP-treated cells.
  • AP did not affect the incorporation or copy number of the H-ras gene, as shown by Southern blot analysis.

Conclusions:

  • Antipain (AP) effectively suppresses H-ras-mediated NIH3T3 cell transformation.
  • The anti-transformation effect of AP is associated with decreased expression of both exogenous H-ras and endogenous c-myc.
  • AP's mechanism of action does not involve alterations in the H-ras gene's integration or copy number.

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