Rapid polymerase chain reaction assay to detect variation in the extent of gene-specific damage between cisplatin- or VP-16-resistant and sensitive lung cancer cell lines.
We previously established a rapid and facile polymerase chain reaction (PCR)-stop assay for quantitation of specific gene damage in very small numbers of cells. The present study investigated whether the PCR-stop assay was able to detect variation in the extent of DNA damage in transcribed active genes between cisplatin- or VP-16-resistant and sensitive cells. The assay demonstrated that about twice as much genetic damage occurs in PC-9 cells than in cisplatin-resistant PC-9/CDDP cells following cisplatin exposure and about 4.6 times more damage occurs in H69 than in VP-16-resistant H69/VP cells following VP-16 exposure. These results show that DNA damage, as detected by PCR-stop assay, correlates with cytotoxicity. In conclusion, the PCR-stop assay could be useful in detecting variation in DNA damage in specific genes.
Oshita
F
F
Division of Medical Oncology, National Cancer Center Hospital, Tokyo.
Saijo
N
N
D001483:Base Sequence; D002945:Cisplatin / Q000494:pharmacology*; D004249:DNA Damage; D004351:Drug Resistance; D005047:Etoposide / Q000494:pharmacology*; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008175:Lung Neoplasms / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D005047:Etoposide; D007041:Hypoxanthine Phosphoribosyltransferase; D002945:Cisplatin