Mutation of the endogenous p53 gene in cells transformed by HPV-16 E7 and EJ c-ras confers a growth advantage involving an autocrine mechanism.
Rat embryo fibroblasts transformed with the HPV-16 E7 gene and the activated c-H-ras gene fall into two distinct phenotypic classes. At high cell density, clones of one class form colonies in methylcellulose supplemented with low serum; at low cell density, these cells display responsiveness to mitogenic factors present in serum-free conditioned medium from rat embryo fibroblasts. In contrast, clones of the second class exhibit an absolute dependency on growth factors present in serum at all cell densities in the methylcellulose colony assay and fail to respond to conditioned medium. We find that the status of the endogenous p53 gene is tightly correlated with these two classes of clones. Clones of the first class contain missense mutations in the p53 gene and have lost the wild-type allele. Clones of the second class express wild-type p53 protein. The importance of mutant p53 expression in reducing the growth factor dependency of transformed clones was confirmed in a separate series of experiments in which rat embryo fibroblasts were transformed with three genes, E7 + ras + mutant p53. The growth behaviour of these triply transfected clones was similar to that of the E7 + ras clones expressing endogenous mutant p53. We demonstrate that the enhanced proliferation of E7 + ras clones expressing mutant p53 protein involves an autocrine mechanism.
Peacock
J W
JW
Ontario Cancer Institute/Princess Margaret Hospital, Toronto, Canada.
Benchimol
S
S
D000818:Animals; D001483:Base Sequence; D002461:Cell Line, Transformed; D002999:Clone Cells; D016895:Culture Media, Serum-Free; D017931:DNA Primers; D004622:Embryo, Mammalian; D005347:Fibroblasts; D005814:Genes, Viral; D016158:Genes, p53; D011905:Genes, ras; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016296:Mutagenesis; D015513:Oncogene Proteins / Q000096:biosynthesis* / Q000235:genetics; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics; D027383:Papillomaviridae / Q000235:genetics*; D050725:Papillomavirus E7 Proteins; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D051381:Rats; D011916:Rats, Inbred F344; D015183:Restriction Mapping; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis
D016895:Culture Media, Serum-Free; D017931:DNA Primers; D015513:Oncogene Proteins; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D016159:Tumor Suppressor Protein p53; C077183:human oncogene protein EJ-ras; C059731:oncogene protein E7, Human papillomavirus type 16