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目的:探讨Anisomycin诱导EAC细胞凋亡的机制。方法:利用MTT法观察Anisomycin对EAC细胞增殖的影响;应用AnnexinV-FITC和PI双染色检测Anisomycin作用下EAC细胞凋亡的变化;琼脂糖凝胶电泳检测Anisomycin作用下EAC细胞的DNA断裂片断;采用RT-PCR检测Anisomycin作用下Caspase-3 mRNA的转录水平;Western Blot分析Anisomycin处理的EAC细胞内Caspase-3蛋白的表达。结果:Anisomycin对EAC细胞增殖的抑制率随浓度的升高而增高,EAC细胞的凋亡水平也上升,随着Anisomycin浓度升高EAC细胞DNA断裂的寡核苷酸片段亦越趋明显,Caspase-3的mRNA和蛋白水平也上调,并明显高于阿霉素组结论:结果表明,Anisomycin能够抑制EAC细胞的增殖,在体外可能通过激活Caspase-3凋亡信号而诱导EAC细胞的凋亡。
Objective: To investigate the mechanism of Anisomycin induced apoptosis in EAC cells. Methods: The effect of Anisomycin on the proliferation of EAC cells was observed by MTT assay. The apoptosis of EAC cells was detected by Annexin V-FITC and PI double staining. The DNA fragmentation of EAC cells was detected by agarose gel electrophoresis. The transcription level of Caspase-3 mRNA by Anisomycin was detected by RT-PCR. The Caspase-3 protein expression in Anisomycin-treated EAC cells was analyzed by Western Blot. Results: The inhibitory rate of Anisomycin on EAC cell proliferation increased with the increase of concentration, and the apoptosis of EAC cells also increased. As the concentration of Anisomycin increased, the DNA fragments of EAC cells became clearer. The expression of Caspase- 3 mRNA and protein levels were also up-regulated, and were significantly higher than the doxorubicin group Conclusion: Anisomycin can inhibit the proliferation of EAC cells in vitro activation of Caspase-3 apoptosis signal may induce apoptosis of EAC cells.