title: Gene-specific damage produced by topoisomerase inhibitors in human lung cancer cells and peripheral mononuclear cells as assayed by polymerase chain reaction-stop assay.
abstract: We have previously reported that the polymerase chain reaction (PCR)-stop assay is an useful technique for investigating gene-specific damage induced by cisplatin, and that the degree of gene-specific damage sustained by peripheral blood mononuclear cells (MNC) when exposed to cisplatin in vitro predicts the response to cisplatin-based chemotherapy. In the current study, we investigated whether PCR-stop assay was suitable for investigating gene- specific damage induced by the topoisomerase I inhibitor CPT-11 or the topoisomerase II inhibitor VP-16, in the human lung cancer cell lines PC-7 and H69, respectively. The cells were incubated with CPT-11 or VP-16 for 24 hours in vitro and the hypoxanthine-phosphoribosyl transferase gene was amplified by PCR. Amplification of a 2.7kb fragment of this gene was clearly inhibited by each drug in a dose dependent manner. The concentration which reduced amplification by 63% (D63, the dose that produces an average of one lesion per single strand of the 2.7kb fragment), was 318 micrograms/ml for PC-7 treated with CPT-11 and 35 micrograms/ml for H69 treated with VP-16. We also used PCR-stop assay to investigate gene-specific damage induced by CPT-11 or VP-16 in adenocarcinoma cells from pleural effusions and MNC from freshly isolated blood obtained from 4 patients with lung cancer. Between-patient variations in the extent of gene-specific damage were observed in both types of cells. These results suggest that PCR-stop assay is suitable for the analysis of interindividual variations in the extent of gene-specific damage induced by topoisomerase inhibitors in human cells.
authors.affiliation: Division of Thoracic Disease, Kanagawa Cancer Center, Yokohama, Japan.
authors.forename: F
authors.initials: F
authors.lastname: Oshita
authors.forename: K
authors.initials: K
authors.lastname: Yamada
authors.forename: I
authors.initials: I
authors.lastname: Nomura
authors.forename: K
authors.initials: K
authors.lastname: Noda
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D002166:Camptothecin / Q000031:analogs & derivatives / Q000494:pharmacology; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D005047:Etoposide / Q000494:pharmacology; D005784:Gene Amplification / Q000187:drug effects*; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D000077146:Irinotecan; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000201:enzymology; D008175:Lung Neoplasms / Q000188:drug therapy / Q000201:enzymology; D016133:Polymerase Chain Reaction / Q000379:methods*; D059004:Topoisomerase I Inhibitors; D014407:Tumor Cells, Cultured / Q000187:drug effects
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D059004:Topoisomerase I Inhibitors; D005047:Etoposide; D000077146:Irinotecan; D007041:Hypoxanthine Phosphoribosyltransferase; D002166:Camptothecin