title: Novel revertants of H-ras oncogene-transformed R6-PKC3 cells.
abstract: Rat 6 fibroblasts that overproduce protein kinase C beta 1 (R6-PKC3 cells) are hypersensitive to complete transformation by the T24 H-ras oncogene; yet T24 H-ras-transformed R6-PKC3 cells are killed when exposed to 12-O-tetradecanoylphorbol-13-acetate (TPA) (W.-L. W. Hsiao, G. M. Housey, M. D. Johnson, and I. B. Weinstein, Mol. Cell. Biol. 9:2641-2647, 1989). Treatment of an R6-PKC3 subclone that harbors a T24 H-ras gene under the control of an inducible mouse metallothionein I promoter with ZnSO4 and TPA is extremely cytocidal. This procedure was used to isolate rare revertants that are resistant to this toxicity. Two revertant lines, R-1a and ER-1-2, continue to express very high levels of protein kinase C enzyme activity but, unlike the parental cells, do not grow in soft agar. Furthermore, these revertants are resistant to the induction of anchorage-independent growth by the v-src, v-H-ras, v-raf, and, in the case of the R-1a line, v-fos oncogenes. Both revertant lines, however, retain the ability to undergo morphological alterations when either treated with TPA or infected with a v-H-ras virus, thus dissociating anchorage independence from morphological transformation. The revertant phenotype of both R-1a and ER-1-2 cells is dominant over the transformed phenotype in somatic cell hybridizations. Interestingly, the revertant lines no longer induce the metallothionein I-T24 H-ras construct or the endogenous metallothionein I and II genes in response to three distinct agents: ZnSO4, TPA, and dexamethasone. The reduction in activity of metallothionein promoters seen in these revertants may reflect defects in signal transduction pathways that control the expression of genes mediating specific effects of protein kinase C and certain oncogenes in cell transformation.
authors.affiliation: Institute of Cancer Research, Columbia University College of Physicians and Surgeons, New York, New York 10032.
authors.forename: R S
authors.initials: RS
authors.lastname: Krauss
authors.forename: S N
authors.initials: SN
authors.lastname: Guadagno
authors.forename: I B
authors.initials: IB
authors.lastname: Weinstein
mesh_terms: D000818:Animals; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D002478:Cells, Cultured; D004789:Enzyme Activation; D015972:Gene Expression Regulation, Neoplastic; D005799:Genes, Dominant; D011905:Genes, ras / Q000235:genetics*; D009024:Morphogenesis; D016296:Mutagenesis; D015513:Oncogene Proteins / Q000096:biosynthesis*; D010641:Phenotype; D011493:Protein Kinase C / Q000235:genetics; D016755:Proto-Oncogene Proteins c-jun / Q000096:biosynthesis; D051381:Rats
chemical_list: D015513:Oncogene Proteins; D016755:Proto-Oncogene Proteins c-jun; D011493:Protein Kinase C