Cell-cycle arrest and p53 accumulation induced by geldanamycin in human ovarian tumour cells.
We have analysed the cell-cycle arrests and cytotoxicity of the A2780 human ovarian cell line in response to geldanamycin, a benzoquinoid ansamycin that can inhibit tyrosine kinases. Geldanamycin causes a dose-dependent G2 arrest and reversible inhibition of entry into the S phase in A2780 cells. After a 3-h exposure to 0.1 microM geldanamycin, the cells show an increase in accumulation of p53 protein that is maximal at 24 h after drug exposure. Increased p53 levels can be induced in cells by DNA-damaging agents; however, using alkaline elution and sister chromatid exchange assays we detect no DNA damage induced by geldanamycin. Using dominant negative mutant TP53 transfectants of A2780 we have analysed the possible dependence of geldanamycin-induced cell-cycle arrests on the presence of functional p53. We observe no difference in cell-cycle arrests in mutant p53 transfectants known to have the p53-DNA damage-response pathway inactivated as compared with vector-alone controls. Similarly, we observe no difference in clonogenic resistance to the cytotoxicity of geldanamycin in these cells. These results suggest that geldanamycin can induce increased p53 protein by a mechanism not involving DNA damage. Furthermore, the cell-cycle arrests and cytotoxic effects of geldanamycin in these cells are not mediated by p53-dependent pathways.
McIlwrath
A J
AJ
CRC Department of Medical Oncology, CRC Beatson Laboratories, Glasgow, UK.
Brunton
V G
VG
Brown
R
R
D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D016227:Benzoquinones; D002453:Cell Cycle / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007399:Interphase / Q000187:drug effects; D047029:Lactams, Macrocyclic; D010051:Ovarian Neoplasms / Q000378:metabolism / Q000473:pathology*; D011809:Quinones / Q000494:pharmacology*; D016196:S Phase / Q000187:drug effects; D012854:Sister Chromatid Exchange; D014162:Transfection; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis*
D000903:Antibiotics, Antineoplastic; D016227:Benzoquinones; D004273:DNA, Neoplasm; D047029:Lactams, Macrocyclic; D011809:Quinones; D016159:Tumor Suppressor Protein p53; C001277:geldanamycin