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[目的]探讨微囊藻毒素(MC)与甲基硝基亚硝基胍(MNNG)对食管癌EC109细胞株毒性的联合作用。[方法]应用MTT法观察微囊藻毒素LR(MC-LR)(1 nmol/L、10 nmol/L、100 nmol/L、1μmol/L和10μmol/L)和MNNG(5 pmol/L、50 pmol/L、500 pmol/L、5 nmol/L、50 nmol/L、500 nmol/L和5μmol/L)对食管癌EC109细胞株生长抑制率(fa)的影响,利用析因设计方差分析和Chou-Talalay联合指数法(中效原理)分析两毒物的联合作用;应用流式细胞术检测细胞凋亡。[结果]MNNG单独染毒,当浓度达到50 pmol/L及以上时,各组fa均明显高于对照组(均P<0.05);MC-LR单独染毒,fa则先降低后增加,除100 nmol/L组外,其余组与对照组相比,差异均有统计学意义(均P<0.05);各浓度MC-LR与MNNG组联合染毒,fa均随着MNNG浓度升高而增加(r值分别为0.961,0.973,0.950,0.980,0.959,0.972,均P<0.01),各浓度MNNG与MC-LR组联合染毒,均随着MC-LR染毒浓度升高fa先降低后升高,与对照组相比差异有统计学意义(P<0.05);析因设计方差分析表明两毒物有交互作用(P<0.01);运用中效原理计算合用指数(CI),当fa=9.4%时,CI=1,两毒物联合呈现相加效应;当fa<9.4%时,CI>1,两毒物联合产生拮抗效应;当fa>9.4%时,CI<1,两毒物联合产生协同效应;流式细胞术检测细胞凋亡结果表明,MNNG与高剂量MC-LR均可诱导细胞凋亡,且高剂量MNNG与MC-LR联合染毒具有协同诱导细胞凋亡的作用。[结论]MC-LR与MNNG联合染毒,低剂量时两者为拮抗效应,高剂量时为协同效应。
[Objective] To investigate the combined effects of microcystin (MC) and methyl nitrosothioguanidine (MNNG) on the toxicity of esophageal carcinoma EC109 cells. [Method] MTT assay was used to observe the effects of microcystin LR (MC-LR) (1 nmol / L, 10 nmol / L, 100 nmol / L, 1 μmol / L and 10 μmol / L) The effects of different concentrations of different concentrations of puerarin on the growth inhibition rate (fa) of esophageal cancer EC109 cells were investigated by means of analysis of variance Chou-Talalay joint index method (the principle of efficiency) to analyze the combined effect of two poisons; apoptosis was detected by flow cytometry. [Results] MNNG alone treated with MC-LR alone could significantly reduce fa and FA fa concentrations at 50 pmol / L and above, and fa of all groups was significantly higher than that of the control group (all P <0.05) (All P <0.05). The concentrations of MNNG increased with the increase of MN concentration in MC-LR and MNNG groups (r values were 0.961,0.973,0.950,0.980,0.959,0.972, respectively, P <0.01), the concentration of MNNG and MC-LR group combined with exposure, with the concentration of MC-LR increased fa first decreased (P <0.05). Analysis of variance by factorial design showed that there was interaction between the two poisons (P <0.01), and CI was calculated by the principle of median efficiency. When fa = At 9.4%, CI = 1, the two poisons showed an additive effect; when fa <9.4%, CI> 1, the two poisons produced an antagonistic effect; when fa> 9.4%, CI <1, The results of apoptosis assay by flow cytometry showed that MNNG and high dose of MC-LR could induce apoptosis, and the combination of high dose MNNG and MC-LR could induce apoptosis. [Conclusion] Both MC-LR and MNNG were antagonistic when exposed to low doses and synergistic at high doses.