Introduction of a murine p53 mutation corresponding to human codon 249 into a murine hepatocyte cell line results in growth advantage, but not in transformation.
The p53 gene is frequently mutated in human tumors; in hepatocellular carcinomas, there is a high frequency of a specific mutation at codon 249 in regions with significant aflatoxin exposure. To assess the role of this p53 mutation in the development of hepatocellular carcinoma, a mutant murine p53 gene, p53ser246, which corresponds to human codon 249, was transfected into a differentiated, nontransformed hepatocyte cell line AML12. Expression of p53ser246 in this line resulted in a growth advantage when compared with either a control vector (which contains a large p53 deletion) or with a different p53 mutant, val135, not found in hepatocellular carcinoma. Overall, there was a threefold increase in colony formation after transfection with p53ser246 as compared with the control or p53val135 vectors, and the p53ser246 plates developed consistently larger colonies. Whereas clones expressing the control or p53val135 constructs showed no significant morphological changes, clones expressing p53ser246 showed increased heterogeneity (large multinucleated cells and areas of small crowded cells) without focus formation. In addition, the ser246 mutation imparted a growth advantage in serum-free media, suggesting less dependence on specific factors present in serum. None of the mutant p53 or control lines were capable of growth in soft agar or tumor formation in nude mice. Thus in this model, in which endogenous wild-type p53 expression is retained, a high level of mutant p53 expression is not sufficient to transform hepatocytes. Our findings indicate that p53ser246 has effects on hepatocytes that may result in a clonal growth advantage and suggest that additional factors are required for the development of hepatocellular carcinoma.
Dumenco
L
L
Department of Pathology and Laboratory Medicine, Brown University, Providence, RI, USA.
Oguey
D
D
Wu
J
J
Messier
N
N
Fausto
N
N
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002455:Cell Division; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D003062:Codon; D015870:Gene Expression; D016158:Genes, p53; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D014162:Transfection
D003062:Codon