In Situ Detection of Cellular DNA Strand Breaks Induced by Antitumor Drugs Using Nick Translation

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Cellular DNA strand breaks induced by an alkylating agent : Carboquone (CQ), and heat (43^ﾟC) was detected in HeLa cells in vitro and mouse sarcoma-180 cells in vivo. The breaks sites in the DNA were translated artificially in the presence of Escherichia coli DNA polymerase I and [ ^3H] -1 abeled dTTp and sites in the DNA were visualized byautoradiographic 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 fordetermining drug and heat induced DNA damage of tumor cells, under in vitro and in vivo conditions and in a semi-quantitative manner.