甾体3-氰基-2-氨基吡啶5g对EC109细胞增殖、凋亡的影响
Effect of steroid 3-cyano-2-aminopyridines compound 5g on proliferation and apoptosis of EC109 cells

目的:考察甾体3-氰基-2-氨基吡啶5g(S3C2A-5g)对食管癌EC109细胞增殖和凋亡的影响。方法:采用不同浓度的S3C2A-5g作用于EC109细胞不同时间后,MTT法检测细胞增殖; 结晶紫染色检测细胞克隆形成; 流式细胞术检测细胞凋亡; DCFH-DA染色检测细胞内活性氧的含量; 采用Hoechst染色和免疫荧光法检测细胞水平的Cleaved Caspase-3表达。结果:S3C2A-5g对EC109细胞的增殖抑制作用有明显的浓度和时间依赖性,0.40 μmol/L S3C2A-5g的抑制率可达到(24.31±1.05)%。S3C2A-5g抑制细胞克隆形成,诱导细胞凋亡。诱导凋亡的机制与活性氧和Cleaved Caspase通路相关。结论:S3C2A-5g通过抑制克隆形成,促进食管癌EC109细胞凋亡,进而抑制细胞增殖。其诱导细胞凋亡的机制可能与细胞内氧化损伤及Caspase家族相关蛋白激活有关。
Aim: To investigate the effect of steroid 3-cyano-2-aminopyridine compound 5g(S3C2A-5g)on proliferation and apoptosis of EC109 cells.Methods: The esophageal cancer cells were treated with different doses of S3C2A-5g for different time. Cell proliferation was detected by MTT assay. Cell clone formation was examined by crystal violet staining. Cells apoptosis was detected by flow cytometry. Intracellular reactive oxygen species levels were detected by DCFH-DA staining. The expression of Cleaved Caspase-3 in the cells was detected by Hoechst staining and immunofluorescence staining.Results: S3C2A-5g could inhibit the proliferation of esophageal cancer EC109 cells concentration and time-dependently. The inhibitory rate of 0.40 μmol/L S3C2A-5g was(24.31±1.05)%. S3C2A-5g could inhibit cell clonal formation and induce apoptosis. The mechanism of inducing apoptosis was related to intracellular reactive oxygen species and Cleaved Caspase pathway.Conclusion: S3C2A-5g can promote the apoptosis of EC109 cells by inhibiting the formation of colonies, and then inhibit the cell proliferation. The mechanism of apoptosis is related to the intracellular oxidative damage and the activation of Caspase family related proteins