Technology Center, China Tobacco Zhejiang Industrial Co. Ltd., Hangzhou 310008, Zhejiang, People's Republic of China.
Mutat Res. 2010 Feb;696(1):48-54. doi: 10.1016/j.mrgentox.2009.12.011. Epub 2009 Dec 24.
In the present study, the cyto-genotoxic effects of 12 CSCs prepared from a diverse set of cigarettes on human B lymphoblastoid cells were compared using five in vitro assays. The cells were exposed to CSCs at doses of 2.5, 5.0, 7.5, 10.0, and 12.5 x 10(-3)cigarette/ml for 24h in neutral red uptake and CCK-8 assays, at doses of 1.0, 2.0, 3.0, 4.0, and 5.0 x 10(-3)cigarette/ml for 3h in cell apoptosis assay, at doses of 6.0, 8.0, 10.0, 12.0, and 14.0 x 10(-3)cigarette/ml for 4h in comet assay, and at doses of 1.0, 2.0, 4.0, 6.0, and 8.0 x 10(-3)cigarette/ml for 4h in micronucleus assay. The potency of 12 CSCs to induce corresponding toxic effects in each assay was calculated, and the correlations between the results in five assays were analyzed. Our investigation showed that the results of 12 CSCs in CCK-8 and cell apoptosis assays were positive, the results of 11 CSCs in neutral red uptake and comet assays were positive, and 9 CSCs could induce significantly the micronuclei in micronucleus assay. It was found that the potency to induce the cytotoxic effects among 12 CSCs ranged 9.694 folds in neutral red uptake assay and 6.43 folds in CCK-8 assay, the potency to induce cell apoptosis among 12 CSCs ranged 8.191 folds, the potency to induce DNA damage among 12 CSCs ranged 29.199 folds, the potency to induce micronuclei among 12 CSCs ranged 5.879 folds. Moreover, the good correlations were found between any two assays. It was suggested that the cyto-genotoxicity of CSCs from different brands of cigarettes varied greatly, comet assay might be a sensitive assay in assessing the genotoxicity induced by CSCs.
在本研究中，使用五种体外检测方法比较了来自不同品牌香烟的 12 种 CSCs 对人 B 淋巴母细胞的细胞遗传毒性作用。将细胞暴露于 CSCs 中，浓度分别为 2.5、5.0、7.5、10.0 和 12.5×10(-3)支香烟/ml 24 小时，用于中性红摄取和 CCK-8 测定；浓度分别为 1.0、2.0、3.0、4.0 和 5.0×10(-3)支香烟/ml 3 小时，用于细胞凋亡测定；浓度分别为 6.0、8.0、10.0、12.0 和 14.0×10(-3)支香烟/ml 4 小时，用于彗星试验；浓度分别为 1.0、2.0、4.0、6.0 和 8.0×10(-3)支香烟/ml 4 小时，用于微核试验。计算了 12 种 CSCs 在每种检测中的诱导相应毒性作用的效力，并分析了五种检测结果之间的相关性。我们的研究表明，12 种 CSCs 在 CCK-8 和细胞凋亡检测中的结果为阳性，11 种 CSCs 在中性红摄取和彗星检测中的结果为阳性，9 种 CSCs 可以在微核检测中显著诱导微核。发现，12 种 CSCs 在中性红摄取检测中诱导细胞毒性作用的效力范围为 9.694 倍，在 CCK-8 检测中为 6.43 倍，12 种 CSCs 诱导细胞凋亡的效力范围为 8.191 倍，12 种 CSCs 诱导 DNA 损伤的效力范围为 29.199 倍，12 种 CSCs 诱导微核的效力范围为 5.879 倍。此外，发现任意两个检测之间都存在良好的相关性。提示不同品牌香烟的 CSCs 细胞遗传毒性差异较大，彗星试验可能是评估 CSCs 诱导遗传毒性的敏感试验。
