Xin Lili, Wang Jianshu, Wu Yanhu, Guo Sifan
School of Public Health, Medical College of Soochow University , Suzhou, Jiangsu , China and.
Toxicol Mech Methods. 2015 Feb;25(2):136-42. doi: 10.3109/15376516.2014.1003357. Epub 2015 Jan 22.
In order to assess the potential carcinogenic and genotoxic responses induced by environmental pollutants, genotoxicity test systems based on a GADD45α promoter-driven luciferase reporter in human A549 and HepG2 cells were established.
Four different types of environmental toxicants including DNA alkylating agents, precarcinogenic agents, DNA cross-linking agents and non-carcinogenic agents, and three environmental samples collected from a coke oven plant were used to evaluate the test systems. After treated with the tested agents and environmental samples for 12 h, the cell viabilities and luciferase activities of the luciferase reporter cells were determined, respectively.
Methyl methanesulfonate, benzo[a]pyrene, formaldehyde and the extractable organic matter (EOM) from coke oven emissions in ambient air generally produced significant induction of relative luciferase activity in a similar dose-dependent manner in A549- and HepG2-luciferase cells. No significant increases in relative luciferase activity were observed in pyrene-treated A549- or HepG2-luciferase cells. Significant increase in relative luciferase activity was already evident after 2.5 µM benzo[a]pyrene, 5 µM formaldehyde, 0.006 µg/L bottom-EOM, 0.10 µg/L side-EOM or 0.06 µg/L top-EOM, where no cytotoxic damage was observed. Compared with the A549-luciferase cells, the tested pollutants produced higher induction of relative luciferase activity in HepG2-luciferase cells.
Therefore, the new genotoxicity test systems can detect different types of genotoxic agents and low concentrations of environmental samples. The luciferase reporter cells, especially the HepG2-luciferase cells, could provide a valuable tool for rapid screening of the genotoxic damage of environmental pollutants and their complex mixtures.
为了评估环境污染物诱导的潜在致癌和遗传毒性反应，建立了基于人A549和HepG2细胞中GADD45α启动子驱动的荧光素酶报告基因的遗传毒性检测系统。
使用四种不同类型的环境毒物，包括DNA烷化剂、致癌前体物质、DNA交联剂和非致癌剂，以及从炼焦厂采集的三个环境样品来评估该检测系统。在用受试物和环境样品处理12小时后，分别测定荧光素酶报告基因细胞的细胞活力和荧光素酶活性。
甲磺酸甲酯、苯并[a]芘、甲醛以及炼焦炉排放的环境空气中的可提取有机物（EOM），通常在A549和HepG2荧光素酶细胞中以类似的剂量依赖性方式显著诱导相对荧光素酶活性。在芘处理的A549或HepG2荧光素酶细胞中未观察到相对荧光素酶活性的显著增加。在2.5μM苯并[a]芘、5μM甲醛、0.006μg/L底部EOM、0.10μg/L侧面EOM或0.06μg/L顶部EOM处理后，相对荧光素酶活性就已显著增加，且未观察到细胞毒性损伤。与A549荧光素酶细胞相比，受试污染物在HepG2荧光素酶细胞中诱导的相对荧光素酶活性更高。
因此，新的遗传毒性检测系统可以检测不同类型的遗传毒性剂和低浓度的环境样品。荧光素酶报告基因细胞，尤其是HepG2荧光素酶细胞，可为快速筛选环境污染物及其复杂混合物的遗传毒性损伤提供有价值的工具。
