TPA-induced differentiation of human rhabdomyosarcoma cells involves dephosphorylation and nuclear accumulation of mutant P53

A. Germani, C. Fusco, S. Martinotti, A. Musaro, M. Molinaro, B. M. Zani

Research output: Contribution to journalArticle

Abstract

Previous studies have shown that human rhabdomyosarcoma cells are induced to differentiate by TPA, in the absence of appreciable alterations of the muscle regulatory genes and their products (1). The question was addressed whether the tumor suppressor p53 could be a target of TPA action in these cells. Genomic analysis by a Polymerase Chain Reaction/Single-Strand Conformation Polymorphism (PCR/SSCP) and direct sequencing indicate the presence of a mutation in exon VII at codon 248 (C to T transition) and a loss of heterozygosity of p53 gene in human rhabdomyosarcoma cell line (RD). It is here shown that transcription of p53 mRNA strongly decreases in RD cells induced to growth arrest and differentiate by TPA treatment. In these cells immunoprecipitation and immunoblot analysis show that both synthesis and total cellular concentration of the protein are also reduced by TPA. Nevertheless nuclear p53 accumulation is at much higher extent, whereas 32 P-orthophosphate labelling, followed by immunoprecipitation, demonstrates a decrease of phosphorylation of both cytoplasmic and nuclear p53. These results indicate that TPA causes a number of alterations of mutant p53, likely mediated through a protein kinase C dependent mechanism, which might impair the transforming ability of mutant p53 in growth-arrested and differentiating RD cells.

Original languageEnglish
Pages (from-to)17-24
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume202
Issue number1
DOIs
Publication statusPublished - 1994

Fingerprint

Rhabdomyosarcoma
Genes
Phosphorylation
Polymerase chain reaction
Transcription
Polymorphism
Labeling
Protein Kinase C
Muscle
Conformations
Tumors
Exons
Immunoprecipitation
Phosphates
Cells
Messenger RNA
Loss of Heterozygosity
p53 Genes
Regulator Genes
Growth

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

TPA-induced differentiation of human rhabdomyosarcoma cells involves dephosphorylation and nuclear accumulation of mutant P53. / Germani, A.; Fusco, C.; Martinotti, S.; Musaro, A.; Molinaro, M.; Zani, B. M.

In: Biochemical and Biophysical Research Communications, Vol. 202, No. 1, 1994, p. 17-24.

Research output: Contribution to journalArticle

Germani, A. ; Fusco, C. ; Martinotti, S. ; Musaro, A. ; Molinaro, M. ; Zani, B. M. / TPA-induced differentiation of human rhabdomyosarcoma cells involves dephosphorylation and nuclear accumulation of mutant P53. In: Biochemical and Biophysical Research Communications. 1994 ; Vol. 202, No. 1. pp. 17-24.
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