Transfection of C3H10T1/2 cells with the Harvey-ras oncogene reduces cytosolic phospholipase A2 function.
Several lines of evidence suggest that phospholipase A2 may play a role in the activated ras-mediated transformation process. In the present study, phospholipase A2 activity and expression were assessed in a murine fibroblast cell line (C3H10T1/2 cells) that was stably transfected with the Harvey ras oncogene, a cellular model used for studying multistage carcinogenesis. Reduced levels of fatty acids were released from the ras-transfected cells compared to untransfected controls. The in vitro phospholipase A2 activity apparent in the C3H10T1/2 showed preference for sn-2 arachidonyl phosphatidylcholine compared to dipalmitoyl phosphatidylcholine. The activity, as well as the cytosolic phospholipase A2 immunoreactive protein, was reduced by 50% in the ras-transfected cells compared to control cells. These results suggest that the cytosolic phospholipase A2 is the predominant form of this enzyme family expressed in C3H10T1/2 cells and that the activity and protein amount is reduced by 50% in these cells when stably transfected with the Harvey ras oncogene.
Department of Foods and Nutrition, Purdue University, West Lafayette, IN 47907-1264, USA.
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Teegarden
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Burgess
D000818:Animals; D002460:Cell Line; D003600:Cytosol / Q000201:enzymology*; D005227:Fatty Acids / Q000378:metabolism; D005347:Fibroblasts / Q000201:enzymology; D011905:Genes, ras / Q000235:genetics*; D051379:Mice; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D013379:Substrate Specificity; D014162:Transfection
D005227:Fatty Acids; D010741:Phospholipases A; D054467:Phospholipases A2