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Transfection of C3H10T1/2 cells with the Harvey-ras oncogene reduces cytosolic phospholipase A2 function

D Teegarden1, X Xu, J R Burgess

  • 1Department of Foods and Nutrition, Purdue University, West Lafayette, IN 47907-1264, USA.

Cancer Letters
|October 1, 1996
PubMed

Insights

Ras oncogene transformation reduces phospholipase A2 (PLA2) activity and protein levels in murine fibroblasts. This suggests PLA2 may be involved in ras-mediated cell transformation and carcinogenesis.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Phospholipase A2 (PLA2) is implicated in cellular transformation processes.
  • The role of PLA2 in ras-mediated transformation requires further investigation.

Purpose of the Study:

  • To assess phospholipase A2 activity and expression in a ras-transfected murine fibroblast cell line.
  • To determine the specific form of PLA2 involved and its modulation by the Harvey ras oncogene.

Main Methods:

  • Utilized a murine fibroblast cell line (C3H10T1/2) stably transfected with the Harvey ras oncogene.
  • Assessed in vitro phospholipase A2 activity using specific phosphatidylcholine substrates.
  • Quantified cytosolic phospholipase A2 protein levels via immunoreactive assays.

Main Results:

  • Ras-transfected cells showed reduced release of fatty acids compared to controls.
  • In vitro PLA2 activity exhibited a preference for sn-2 arachidonyl phosphatidylcholine.
  • Both PLA2 activity and cytosolic PLA2 protein levels were decreased by approximately 50% in ras-transfected cells.

Conclusions:

  • Cytosolic phospholipase A2 is the predominant form in C3H10T1/2 cells.
  • Harvey ras oncogene transfection significantly reduces both cytosolic PLA2 activity and protein expression.
  • These findings suggest a role for reduced PLA2 in ras-mediated cellular transformation and multistage carcinogenesis.

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