Maehara Y, Anai H, Kusumoto T, Sakaguchi Y, Emi Y, Kohnoe S, Sugimachi K
Second Department of Surgery, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Nihon Geka Gakkai Zasshi. 1989 Sep;90(9):1576-8.
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.
在体外培养的HeLa细胞和体内的小鼠肉瘤-180细胞中，检测到了由烷化剂卡波醌（CQ）和热（43℃）诱导的细胞DNA链断裂。在大肠杆菌DNA聚合酶I和[³H]标记的dTTP存在的情况下，人工翻译DNA中的断裂位点，并通过对细胞核中银粒的放射自显影观察来可视化DNA中的位点。与对照细胞的结果相比，这些断裂以剂量和时间依赖性方式增加。我们的研究结果表明，暴露于CQ或热后，细胞的存活反应降低，而DNA链断裂水平增加。缺口平移法是一种在体外和体内条件下以半定量方式测定药物和热诱导的肿瘤细胞DNA损伤的快速原位检测方法。
