Nucleotide excision repair in the human ovarian carcinoma cell line (2008) and its cisplatin-resistant variant (C13*).
Repair of cisplatin-damaged DNA was investigated in a human ovarian carcinoma cell line (2008) and its cisplatin-resistant variant (C13*) using a host-cell reactivation (HCR) assay. The HCR of cisplatin-damaged adenovirus (Ad) was not significantly different in C13* cells compared to 2008 cells. The cisplatin concentrations required to reduce the amount of viral DNA replicated to 50% were 0.12 +/- 0.02 microM and 0.10 +/- 0.01 microM after 48 h of repair in 2008 and C13* cells respectively. Similarly, the cisplatin concentration required to reduce the expression of a reporter gene inserted in the viral DNA was not significantly altered in C13* cells compared to the parental line (IC50 values were 0.28 +/- 0.04 microM in 2008 cells and 0.17 +/- 0.06 microM in C13* cells after 48 h of repair). Pretreatment of the cells with cisplatin, immediately prior to Ad infection, did not significantly alter the HCR of cisplatin-damaged Ad in either cell type. In addition, a cisplatin-sensitive variant derived from the C13* cells, namely the RH4 cells, did not differ significantly from either the 2008 or C13* cells in their ability to reactivate cisplatin-damaged Ad. Furthermore, a component of the nucleotide excision repair (NER) pathway, DNA polymerase alpha, was investigated using the competitive inhibitor aphidicolin. The combination of cisplatin and aphidicolin resulted in similar synergistic growth inhibition in both the 2008 and C13* cells providing additional support to the HCR results which suggest that enhanced NER is not responsible for the cisplatin resistance in C13* cells.
Moorehead
R A
RA
Department of Pathology, McMaster University, Hamilton, Ontario, Canada.
Armstrong
S G
SG
Rainbow
A J
AJ
Singh
G
G
D000256:Adenoviridae / Q000187:drug effects / Q000235:genetics; D000970:Antineoplastic Agents / Q000633:toxicity*; D016590:Aphidicolin / Q000633:toxicity; D002277:Carcinoma / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000633:toxicity*; D018736:DNA Adducts / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects*; D004257:DNA Polymerase II / Q000378:metabolism; D004260:DNA Repair; D004279:DNA, Viral / Q000187:drug effects; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000633:toxicity; D005260:Female; D006801:Humans; D010051:Ovarian Neoplasms / Q000473:pathology*; D014407:Tumor Cells, Cultured / Q000187:drug effects
D000970:Antineoplastic Agents; D018736:DNA Adducts; D004279:DNA, Viral; D004791:Enzyme Inhibitors; D016590:Aphidicolin; D004257:DNA Polymerase II; D002945:Cisplatin