Detection of cellular DNA strand breaks induced by antitumor drug and heat using in situ nick translation
Maehara, Y.; Anai, H.; Kusumoto, T.; Sakaguchi, Y.; Emi, Y.; Kohnoe, S.; Sugimachi, K.
Nihon Geka Gakkai Zasshi 90(9): 1576-1578
1989
ISSN/ISBN: 0301-4894 PMID: 2586469 Document Number: 346233
Cellular DNA strand break induced by an alkylating agent: Carboquone (CQ), and heat (43 degrees C) was detected in HeLa cells in vitro and mouse sarcoma-180 cells in vivo. The break sites in the DNA were translated artificially in the presence of Escherichia coli DNA polymerase I and [3H]-labeled dTTP and sites in the DNA were visualized by autoradiographic observation of grains in the nuclei. These breaks increased in a dose and time dependent manner, compared to findings in the control cells. Our findings show that the surviving response of cells decreases while the level of DNA strand breaks increases following exposure to CQ or heat. The nick translation method is a rapid in situ assay for determining drug and heat induced DNA damage of tumor cells, under in vitro and in vivo conditions and in a semi-quantitative manner.