Inhibition of the mutant p53 gene in transformation assays

Samuel W. Beenken, Loretta Raycroft, Guillermina Lozano

Research output: Contribution to journalArticle

Abstract

Mutation of the p53 gene is a key element in the development of several human cancers. Intron 4, a noncoding region of the p53 gene, is required for optimal expression of that gene. We have previously shown that nuclear protein binds intron 4 and have defined the protein-binding site. In this paper we address the question, "Does the mutant p53 gene's ability to transform cells to the malignant phenotype depend on protein binding to intron 4?" Using an in vitro assay in which the mutant p53 gene and Ha-ras oncogene cooperate in transformation of cells to the malignant phenotype, we determined the ability of mutant mouse p53 gene constructs, with and without two base pair substitutions at the intron 4 protein-binding site, to participate in malignant transformation. On Day 1, 5 × 105 rat embryo fibroblasts were transfected by the calcium phosphate procedure with 10 μg of both a mutant p53 gene construct and Ha-ras oncogene. Malignant transformation was evidenced by the formation of discrete foci of heaped-up cells. After 14 days of incubation at 37°C in DMEM and 10% fetal calf serum (8% CO2), the cells were stained with cresyl violet and the foci counted. In three separate experiments, the presence of two base pair substitutions at the intron 4 protein-binding site caused a significant decrease in the number of foci formed (P < 0.05).

Original languageEnglish (US)
Pages (from-to)401-405
Number of pages5
JournalJournal of Surgical Research
Volume52
Issue number4
DOIs
StatePublished - Apr 1992

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p53 Genes
Protein Binding
Introns
Aptitude
ras Genes
Binding Sites
Base Pairing
Inteins
Phenotype
Human Development
Nuclear Proteins
Inhibition (Psychology)
Embryonic Structures
Fibroblasts
Gene Expression
Mutation
Serum
Neoplasms

ASJC Scopus subject areas

  • Surgery

Cite this

Inhibition of the mutant p53 gene in transformation assays. / Beenken, Samuel W.; Raycroft, Loretta; Lozano, Guillermina.

In: Journal of Surgical Research, Vol. 52, No. 4, 04.1992, p. 401-405.

Research output: Contribution to journalArticle

Beenken, Samuel W. ; Raycroft, Loretta ; Lozano, Guillermina. / Inhibition of the mutant p53 gene in transformation assays. In: Journal of Surgical Research. 1992 ; Vol. 52, No. 4. pp. 401-405.
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