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An.nexinV—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-3mRNA的转录水平;WesternBlot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 effects of Anisomycin on apoptosis of ehrlich ascites carcinoma (EAC). Methods: In vitro model of EAC cells was used to detect the effects of Anisomycin on the proliferation of EAC cells through MTT assay. The effects of Anisomycin on apoptosis of EAC cells were detected by AnnexinV-FITC/PI double staining assay. RT-PCR was used to determine the level of Caspase-3 mRNA in EAC cells treated with Anisomycin. Western Blotting assay was applied to analyze the expression of Caspase-3 of EAC cells treated by Anisomycin. Results: The inhibition rate of Anisomycin on EAC cell proliferation increased with the increasing of Anisomycin concentration. The apoptosis rate of Anisomycin-treated EAC cells detected by flow cytometry increased with the increasing of Anisomycin concentration. RT-PCR showed that the level of Caspase-3 mRNA in EAC cells increased with the increasing of Anisomycin. Western blotting showed that the level of Caspase-3 expression in EAC cells increased with the increasing of Anisomycin. Conclusions: These results indicate that Anisomycin can inhibit the EAC cell growth and promote the apoptosis of EAC cells by activating the Caspase-3 signaling