Analysis of phospholipid metabolism in murine keratinocytes transformed by the v-ras oncogene: Relationship of phosphatidylinositol turnover and cytokine stimulation to the transformed phenotype

Edmund Lee, Kari Punnonen, Christina Cheng, Adam Click, Andrzej Dlugosz, Stuart H. Yuspa

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

Introduction of a v-rasHa oncogene into cultured mouse keratinocytes by transduction with a defective retrovirus is sufficient to transform keratinocytes to the benign phenotype. Transduced keratinocytes overexpress TGFα and hyperproliferate in culture medium with 0.05 mM Ca2+. Whereas normal keratinocytes respond to elevated medium Ca2+ by cessation of proliferation and induction of terminal differentiation, v-rasHa keratinocytes are not induced to differentiate by Ca2+. We now demonstrate that v-rasHa keratinocytes have elevated basal levels of phosphatidylinositol, inositol phosphates and diacylglycerols in 0.05 mM Ca2+ medium. Basal turnover of phosphatidylcholine is not altered by the rasHa oncogene. The generation of inositol phosphates is even further stimulated in v-rasHa cells by an increase in extracellular Ca2+ or by exposure to aluminum fluoride. Thus, the v-rasHa gene product does not stimulate the inositol phospholipid pathway maximally and additional phosphatidylinositol is available for turnover in response to inducers of phospholipase C activity. TGFα and medium conditioned by v-rasHa keratinocytes, both of which stimulate proliferation of normal cells in 0.05 mM Ca2+, transiently increased phosphatidylinositol turnover in normal keratinocytes but did not inhibit Ca2+-induced terminal differentiation. In contrast, sustained elevation in basal phosphatidylinositol metabolism was produced by aluminum fluoride. Combined exposure to aluminum fluoride and exogenous TGFα caused hyperproliferation, resistance to Ca2+-induced differentiation and morphological changes identical to those of v-rasHa keratinocytes. These results provide a link between the biological consequences of v-rasHa gene expression and biochemical changes which are known to alter the keratinocyte phenotype.

Original languageEnglish (US)
Pages (from-to)2367-2373
Number of pages7
JournalCarcinogenesis
Volume13
Issue number12
DOIs
StatePublished - Dec 1992

All Science Journal Classification (ASJC) codes

  • Cancer Research

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