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以体外培养人Bel-7402肝癌细胞为模型,研究铅的3种常见化合物氯化铅(Pb Cl2)、乙酸铅(Pb(CH3COO)2)、硝酸铅(Pb(NO3)2)的细胞毒性和去甲基化表观遗传毒性。应用MTS方法检测细胞的存活率,以前期研究建立的评价方法评价铅化合物的去甲基化表观遗传毒性。结果显示,Pb Cl2、Pb(CH3COO)2、Pb(NO3)2均会抑制Bel-7402细胞的增值,计算求得Pb Cl2、Pb(CH3COO)2、Pb(NO3)2相应的50%细胞存活浓度(IC50)值分别为2 524μmol·L~(-1)、1 977μmol·L~(-1)、1 899μmol·L~(-1);80%细胞存活浓度(IC80)值分别为264μmol·L~(-1)、221μmol·L~(-1)、281μmol·L~(-1),通过对3种染毒物不同染毒浓度的细胞存活率进行随机区组设计的方差分析显示3种化合物间的差异无统计学意义(F=0.11;P=0.897)。去甲基化表观遗传毒性检测结果显示,Pb Cl2、Pb(CH3COO)2、Pb(NO3)2均可观察到明显的去甲基化表观遗传毒性,其相对于5-氮杂-2-脱氧胞苷(5-Aza-Cd R)的去甲基化表观遗传毒性当量分别为2.82E-03、1.50E-03、5.09E-04,三者间也无显著性差异。结果表明,铅化合物会使Bel-7402细胞的细胞存活率和转染进细胞的质粒上增强型绿色荧光蛋白基因启动子的DNA甲基化水平下降。
To investigate the cytotoxicity of PbCl2, Pb (CH3COO) 2 and Pb (NO3) 2) in vitro and in vitro of Bel-7402 hepatocarcinoma cells. Demethylation Epigenetics. MTS method was used to detect cell viability. The demethylation epigenetic toxicity of lead compounds was evaluated by the evaluation method established in the previous study. The results showed that Pb Cl2, Pb (CH3COO) 2 and Pb (NO3) 2 all inhibited the proliferation of Bel-7402 cells, and the corresponding 50% of the cells of Pb Cl2, Pb (CH3COO) 2 and Pb The IC50 values were 2 524μmol·L -1, 1 977μmol·L -1 and 1 899μmol·L -1, respectively. The IC50 values of 80% cell were 264μmol · L -1, L -1, 221μmol·L -1, 281μmol·L -1. The variance analysis of the randomized block design of the cell survival rates of three kinds of toxins exposed to different concentrations showed that three kinds The difference between the compounds was not statistically significant (F = 0.11; P = 0.897). The results of demethylation epigenetic toxicity test showed that obvious demethylation epigenetic toxicity was observed in Pb Cl2, Pb (CH3COO) 2 and Pb (NO3) 2, - deoxycytidine (5-Aza-Cd R) demethylation epigenetic equivalent toxicity were 2.82E-03,1.50E-03,5.09E-04, there was no significant difference between the three. The results showed that the lead compound decreased the cell viability of Bel-7402 cells and the DNA methylation level of the enhanced green fluorescent protein gene promoter in the transfected cells.