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致癌物处理后,通过液体闪烁计数法测量HeLa细胞中“非预定”DNA合成。

Measurement of 'unscheduled' DNA synthesis in HeLa cells by liquid scintillation counting after carcinogen treatment.

作者信息

Martin C N, McDermid A C, Garner R C

出版信息

Cancer Lett. 1977 May;2(6):355-60. doi: 10.1016/s0304-3835(77)80017-7.

Abstract

HeLa cells, conditioned in an arginine-deficient medium to reduce DNA S-phase synthesis, were treated with one of four ultimate carcinogens (MNNG, BrMBA, N-acetoxy AAF and EMS) and one precarcinogen, AFB1. All treated cells preferentially incorporated [3H] thymidine as a result of DNA repair monitored by liquid scintillation counting of the extracted DNA. The cells showed some capacity to activate AFB1, but repair synthesis was much increased if a rat liver mixed function oxidase preparation was also present. At equimolar concentrations the various carcinogens stimulated different amounts of DNA repair; this variation was not proportional to the carcinogenic potency of the chemicals tested. Reasons for this are discussed as is the use of this technique as a screen for chemical carcinogens.

摘要

将HeLa细胞置于精氨酸缺乏的培养基中培养,以减少DNA S期合成,然后用四种最终致癌物(MNNG、BrMBA、N-乙酰氧基AAF和EMS)之一以及一种前致癌物AFB1进行处理。通过对提取的DNA进行液体闪烁计数监测DNA修复情况,结果发现所有处理过的细胞都优先掺入了[3H]胸苷。这些细胞表现出一定的激活AFB1的能力,但如果同时存在大鼠肝脏混合功能氧化酶制剂,则修复合成会大大增加。在等摩尔浓度下,各种致癌物刺激的DNA修复量不同;这种差异与所测试化学物质的致癌效力不成正比。文中讨论了出现这种情况的原因以及将该技术用作化学致癌物筛选方法的情况。

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