Technology Center, China Tobacco Zhejiang Industrial Co. Ltd, Hangzhou 310008, Zhejiang, PR China.
Mutat Res. 2010 Mar 29;697(1-2):55-9. doi: 10.1016/j.mrgentox.2010.01.014. Epub 2010 Feb 6.
The differences of the cytogenotoxicity and proteins expression of human B-cell lymphoblastoid 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 lymphoblastoid cell line was exposed to CSCs from two cigarettes (which delivers approximately 3mg tar, 0.3mg nicotine, 3mg CO per cigarette for cigarette 1 and 15mg tar, 1.3mg nicotine, 15mg CO per cigarette for cigarette 2), and the exposure doses were 2.5, 5.0, 7.5, 10.0 and 12.5x10(-3)cigarettes/ml of CSCs for 24h in CCK-8 assay, 6.0, 8.0, 10.0, 12.0 and 14.0x10(-3)cigarettes/ml of CSCs for 4h in comet assay, and 10.0x10(-3)cigarettes/ml of CSCs for 4h 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 p57(Kip2) Ab-7) 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.
采用 CCK-8 法、彗星试验、蛋白质芯片和 Western blot 法检测两种香烟烟雾凝聚物（CSCs）对人 B 淋巴细胞白血病细胞系的体外细胞遗传毒性和蛋白表达的差异。将人 B 淋巴细胞白血病细胞系暴露于两种香烟的 CSCs（香烟 1 每支产生约 3mg 焦油、0.3mg 尼古丁、3mg CO，香烟 2 每支产生 15mg 焦油、1.3mg 尼古丁、15mg CO）中，CCK-8 法中暴露剂量为 2.5、5.0、7.5、10.0 和 12.5x10(-3)支香烟/ml 的 CSCs 24h，彗星试验中暴露剂量为 6.0、8.0、10.0、12.0 和 14.0x10(-3)支香烟/ml 的 CSCs 4h，蛋白水平分析中暴露剂量为 10.0x10(-3)支香烟/ml 的 CSCs 4h。本研究的 CCK-8 法和彗星试验结果表明，香烟 2 组的细胞遗传毒性明显高于香烟 1 组。蛋白质芯片和 Western blot 试验结果表明，香烟 1 组和香烟 2 组之间有 4 种蛋白（即 RAR-beta、14-3-3 sigma、XPF 和 p57(Kip2) Ab-7）的表达水平存在差异。因此，RAR-beta、14-3-3 sigma、XPF 和 p57(Kip2) Ab-7 蛋白可能作为研究 CSCs 诱导细胞遗传毒性的分子生物标志物。
