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目的:观察小分子甾族多取代苯胺化合物2,6-二氰基-3-吡啶基-5-雄甾烯基苯胺(简称4d)对食管癌EC109细胞增殖、周期、凋亡的影响.方法:MTT法检测0.40～100.00 μmol/L的4d作用于正常的食管上皮细胞Her-1a或食管癌细胞EC109 48 h或72 h后,对正常细胞及癌细胞的增殖抑制作用,筛选出4d对食管癌细胞最佳作用时间及浓度;4d作用24 h后,采用流式细胞术检测细胞凋亡及细胞周期;4d作用6h后,使用JC-1染料染色,采用高通量筛选系统检测细胞线粒体膜电位;4d作用24 h后,采用Western blot法检测Bel-2家族中Bcl-2、Mcl-1、Bax蛋白表达的变化.结果:4.0 μmol/L 4d能引起EC109细胞凋亡率升高,S期阻滞,线粒体功能障碍,Mcl-1表达下降(P＜0.05).结论:4d能抑制EC109细胞增殖,诱导细胞凋亡,引起S期阻滞;其机制可能与Bel-2凋亡家族介导的线粒体功能障碍有关.“,”Aim:To investigate the influence of small molecule steroidal polysubstituted aniline compound 2,6-dicyano-3-pyridyl-5-androstenyl aniline(4d) on the proliferation,cell cycle and apoptosis of EC109 cells.Methods:MTT assay was used to detect the proliferation of Het-1 a cells and EC109 cells treated with different concentrations (0.40-100.00 μmoL/L) of 4d for 48 h or 72 h,and the optimal time and concentration of 4d on EC109 cells were screened.After 4d treatment for 24 h,flow cytometry was used to detect the apoptosis rate and cell cycle.After 4d treatment for 6 h,the mitochondrial membrane potential was detected by JC-1 dye staining and high content screening system.After 4d treatment for 24 h,the expressions of Bcl-2,Mcl-1 and Bax in the Bcl-2 family were detected by Western blot.Results:4d at 4.0 μmol/L could cause EC109 cell apoptosis,S cycle block,mitochondrial dysfunction as well as inhibiting the expression of Mcl-1 (P ＜ 0.05).Conclusion:4d could inhibit EC109 cell proliferation,induce apoptosis,and lead to the stagnation of S cycle,which may be associated with Bcl-2 apoptotic family-mediated mitochondrial dysfunction.