Comparison between two kinds of cigarette smoke condensates (CSCs) of the cytogenotoxicity and protein expression in a human B-cell lymphoblastoid cell line using CCK-8 assay, comet assay and protein microarray
Comparison between two kinds of cigarette smoke condensates (CSCs) of the cytogenotoxicity and protein expression in a human B-cell lymphoblastoid cell line using CCK-8 assay, comet assay and protein microarray
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DOI：
10.1016/j.mrgentox.2010.01.014
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发表时间：
2010-03-29
影响因子：
1.9
通讯作者：
He Jiliang
中科院分区：
文献类型：
--
作者：
Lou Jianlin;Chu Guohai;He Jiliang
采用CCK-8法、彗星法、蛋白质微阵列法和蛋白质印迹法体外检测两种香烟烟雾冷凝物(CSC)对人B细胞淋巴母细胞的细胞遗传毒性和蛋白表达的差异。人 B 细胞淋巴母细胞系暴露于两支香烟的 CSC（香烟 1 每支香烟输送约 3 mg 焦油、0.3 mg 尼古丁、3 mg CO，香烟 2 每支香烟输送约 15 mg 焦油、1.3 mg 尼古丁、15 mg CO），暴露接近值为 2.5、5.0、7.5、10.0 和 12.5 x在 CCK-8 测定中，10(-3) 支香烟/ml CSCs，持续 24 小时；在彗星测定中，411 中，6.0、8.0、10.0、12.0 和 14.0 x 10(-3) 支香烟/ml CSCs；在蛋白质水平分析中，10.0 x 10-3 支香烟/ml CSCs，持续 4 小时。本研究中的CCK-8试验和彗星试验结果表明，卷烟2组的细胞遗传毒性显着高于卷烟1组。蛋白质微阵列和Western blot检测结果显示，卷烟1组和卷烟2组四种蛋白（RAR-β、14-3-3 sigma、XPF和p52（Kip2））的表达水平存在差异。因此，RAR-β、14-3-3 sigma、XPF 和 p57(Kip2) Ab-7 蛋白可能作为研究 CSC 诱导的细胞遗传毒性的分子生物标志物。 (C) 2010 Elsevier B.V. 保留所有权利。
The differences of the cytogenotoxicity and proteins expression of human B-cell lymphoblastoicl cells exposed to cigarette smoke condensates (CSCs) from two kinds of cigarettes were detected with CCK-8 assay, comet assay, protein microarray and western blot assay in vitro. Human B-cell lymphoblastoicl cell line was exposed to CSCs from two cigarettes (which delivers approximately 3 mg tar, 0.3 mg nicotine, 3 mg CO per cigarette for cigarette 1 and 15 mg tar, 1.3 mg nicotine, 15 mg CO per cigarette for cigarette 2), and the exposure closes were 2.5, 5.0, 7.5, 10.0 and 12.5 x 10(-3) cigarettes/ml of CSCs for 24 h in CCK-8 assay, 6.0, 8.0, 10.0, 12.0 and 14.0 x 10(-3) cigarettes/ml of CSCs for 411 in comet assay, and 10.0 x 10-3 cigarettes/ml of CSCs for 4 h in protein levels analysis. The results of CCK-8 assay and comet assay in the present study suggested that the cytogenotoxicity in cigarette 2 group was significantly higher than that in cigarette 1 group. The results of protein microarray and western blot assay showed that there were the differences of the expression levels of four proteins (i.e., RAR-beta, 14-3-3 sigma, XPF, and p52(Kip2)) between cigarette 1 group and cigarette 2 group. Hence, it is possible that the RAR-beta, 14-3-3 sigma, XPF, and p57(Kip2) Ab-7 proteins serve as the molecular biomarkers in studying the cytogenotoxicity induced by CSCs. (C) 2010 Elsevier B.V. All rights reserved.