Radiation-induced cell cycle delays and p53 status of early passage melanoma cell lines.
Cell cultures exposed to DNA-damaging agents such as gamma radiation respond by arresting at cell cycle checkpoints, and the p53 tumor suppressor protein is strongly implicated in this behavior. We have investigated the TP53 status and cell cycle response to ionizing radiation of a series of early passage cell lines (designated NZM1 to NZM15) previously developed from patients with metastatic melanoma. The TP53 status of each of the cell lines was determined by single-strand conformation polymorphism and DNA sequence analysis. The majority of the lines appeared to have a wild-type TP53 gene sequence, consistent with published studies. Two lines (NZM4 and NZM7.2) were found to have an identical T-->C transition mutation in nucleotide 721 (exon 7) of the coding region. NZM7.2 (mutant) and NZM7.4 (wild-type) were clonally derived from the same line (NZM7). The existence of radiation-induced cell cycle arrest in G and/or G2M phase was determined 16 h after irradiation (6.3 Gy) by DNA staining and flow cytometric analysis. The mitotic inhibitor paclitaxel was used as a reference compound, with or without irradiation, to assess the efficiency of radiation-induced cell cycle arrest. G1 phase arrest was associated only with the presence of the wild-type TP53 gene, but the efficiency of induced arrest varied among the cell lines and the period of G phase arrest appeared to be short. A significant difference (P < 0.002) was also found between the efficiency of induction of G2 phase arrest and the presence of wild-type TP53 gene. The results provide evidence that although the melanoma cell lines generally had an intact TP53 gene, the efficiency of p53-mediated cycle arrest might be deficient and contribute to the resistance of this tumor to treatment.
Parmar
J
J
Auckland Cancer Society Research Centre, University of Auckland, New Zealand.
Marshall
E S
ES
Charters
G A
GA
Holdaway
K M
KM
Shelling
A N
AN
Baguley
B C
BC
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002453:Cell Cycle / Q000528:radiation effects*; D004273:DNA, Neoplasm / Q000737:chemistry; D005260:Female; D005434:Flow Cytometry; D016158:Genes, p53; D006801:Humans; D008297:Male; D008545:Melanoma / Q000209:etiology / Q000235:genetics* / Q000473:pathology; D008875:Middle Aged; D018807:Polymorphism, Single-Stranded Conformational; D013936:Thymidine / Q000378:metabolism; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D013936:Thymidine