Identification of p53 genetic suppressor elements which confer resistance to cisplatin.
Loss of p53 function is associated with the acquisition of cisplatin resistance in the human ovarian adenocarcinoma A2780 cell line. Selection for cisplatin resistance of A2780 cells was used to isolate genetic suppressor elements (GSEs) from a retroviral library expressing random fragments of human or murine TP53 cDNA. Six GSEs were identified, encoding either dominant negative mutant peptides or antisense RNA molecules which corresponded to various regions within the TP53 gene. Both types of GSE induced cisplatin resistance when introduced individually into A2780 cells. Expression of antisense GSEs led to decreased intracellular levels of p53 protein. One sense GSE induced loss of p53-mediated activities such as DNA damage induced cell cycle arrest and apoptosis. A synthetic peptide, representing the predicted amino acid sequence of this GSE, conferred resistance to cisplatin when introduced into A2780 cells and inhibited the sequence specific DNA binding activity of p53 protein in vitro. Overall, these results directly indicate that inactivation of p53 function confers cisplatin resistance in these human ovarian tumour cells. We have identified short structural domains of p53 which are capable of independent functional interactions and highlighted the efficacy of this approach to discriminate biologically active GSEs from a random fragment library.
Gallagher
W M
WM
CRC Department of Medical Oncology, CRC Beatson Laboratories, Glasgow, UK.
Cairney
M
M
Schott
B
B
Roninson
I B
IB
Brown
R
R
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D016375:Antisense Elements (Genetics) / Q000235:genetics* / Q000302:isolation & purification; D002945:Cisplatin / Q000494:pharmacology*; D004249:DNA Damage / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D005260:Female; D016158:Genes, p53 / Q000235:genetics*; D006801:Humans; D051379:Mice; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D012697:Serine Endopeptidases / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
D000970:Antineoplastic Agents; D016375:Antisense Elements (Genetics); D018076:DNA, Complementary; D016159:Tumor Suppressor Protein p53; C097082:Pep1 protein, mouse; D012697:Serine Endopeptidases; D002945:Cisplatin