Interlaboratory validation of a new assay for DNA-protein crosslinks
Interlaboratory validation of a new assay for DNA-protein crosslinks
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DOI：
10.1016/s0165-1218(96)90043-9
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发表时间：
1996-07-10
期刊：
影响因子：
--
通讯作者：
Bogdanffy, M
中科院分区：
文献类型：
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作者：
Costa, M;Zhitkovich, A;Bogdanffy, M
1992年，建立了一种简便、灵敏的dna -蛋白交联检测方法。为了促进该分析的更广泛使用，进行了一些研究以评估其可靠性和可重复性。在这项工作中，该试验用于评估各种金属和其他化合物是否可以在培养的人淋巴细胞（爱泼斯坦-巴尔病毒转化的伯基特淋巴瘤细胞系）中诱导交联。高锰酸钾、氯化汞、硝酸铅、高氯酸镁、氯化铝和氯化镉在细胞毒性或非细胞毒性水平下均不诱导dna -蛋白质交联，硫酸铜、三氧化二砷和铬酸钾仅在细胞毒性浓度下诱导dna -蛋白质交联。在接触金属化合物5天后，通过细胞增殖和台盼蓝排斥反应评估细胞的急性致死率，通过长期致死率评估细胞的长期致死率。将培养的人淋巴细胞分别用不同剂量的醋酸铅、氯化镉、三氧化二砷和硫酸铜以及顺铂和铬酸盐处理，送到4个不同的实验室，比较dna -蛋白交联试验的可靠性和可重复性。根据所研究的化学物质的不同，在使用该测定法的各个实验室中观察到的结果存在定量差异。然而，所有实验室普遍表明，顺铂、铬酸盐、三氧化二砷和硫酸铜诱导的dna -蛋白质交联水平会产生急性细胞毒性，而氯化镉和醋酸铅则不会。
In 1992, a simple and sensitive assay for detecting DNA-protein crosslinks was developed [1]. In an effort to facilitate the greater use of the assay, a number of studies were conducted to evaluate its reliability and reproducibility. During this work, the assay was used to assess whether various metals and other compounds could induce crosslinks in cultured human lymphocytes (Epstein-Barr virus-transformed Burkitt's Lymphoma cell line). Potassium permanganate, mercury chloride, lead nitrate, magnesium perchlorate, aluminum chloride, and cadmium chloride did not induce DNA-protein crosslinks at either cytotoxic or non-cytotoxic levels, Copper sulfate, arsenic trioxide, and potassium chromate induced DNA-protein crosslinks only at cytotoxic concentrations. Acute lethality of the cells was assessed immediately after exposure to metals by trypan blue exclusion while long-term lethality was assessed by cell proliferation and trypan blue exclusion following an incubation period of 5 days after exposure to the metal compound. All metals exhibited more toxicity in the long-term lethality assay compared to the short-term assay, The cultured human lymphocytes treated with various doses of lead acetate, cadmium chloride, arsenic trioxide and copper sulfate, as well as cis-platinum and chromate, were sent to four different laboratories to compare the reliability and reproducibility of the DNA-protein crosslink assay. Depending on the chemical studied, there were quantitative differences in the results observed among the various laboratories using the assay. However, all laboratories generally showed that cis-platinum, chromate, arsenic trioxide and copper sulfate induced DNA-protein crosslinks at levels that produced acute cytotoxicity, whereas cadmium chloride and lead acetate did not.