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目的探讨苦瓜籽核糖体失活蛋白MAP30及其聚乙二醇修饰物对人肺腺癌细胞A549增殖、细胞凋亡的影响和caspase3相关凋亡途径的诱导作用。方法用MTT法测定MAP30和MAP30-PEG结合物对人肺腺癌细胞A549的抑制率;通过Hoechst33258荧光染色观察A549细胞凋亡形态,并用流式细胞术测定细胞凋亡率以及用western blot检测凋亡细胞caspase3表达。结果 MAP30和MAP30-PEG结合物均对A549有抑制作用,且呈时间和剂量依赖性关系;MAP30-PEG结合物对A549细胞增值抑制率约为MAP30的60%;Hoechst33258荧光染色后均呈典型凋亡形态;流式细胞术检测显示,MAP30和MAP30-PEG结合物作用于A549细胞48 h后均可使A549细胞呈现Sub-G1凋亡峰,凋亡率分别为58.7%、42.7%;MAP30和MAP30-PEG结合物对胞内caspase3在凋亡过程中显示有激活作用。结论 MAP30和MAP30-PEG结合物均对A549增殖有明显的抑制作用;两者均能通过caspase3途径诱导细胞凋亡。
Objective To investigate the effects of bitter melon seed ribosome inactivating protein MAP30 and its modified polyethylene glycol on the proliferation and apoptosis of human lung adenocarcinoma A549 cells and the induction of caspase3-related apoptotic pathway. Methods The inhibitory rates of MAP30 and MAP30-PEG conjugate on human lung adenocarcinoma A549 cells were determined by MTT assay. The apoptosis of A549 cells was observed by Hoechst33258 staining. The apoptosis rate of A549 cells was determined by flow cytometry and Western blot Death cells caspase3 expression. Results MAP30 and MAP30-PEG conjugates all inhibited A549 cells in a time-and dose-dependent manner. MAP30-PEG conjugates inhibited the proliferation of A549 cells by about 60% of that of MAP30 cells. Hoechst33258 cells showed typical apoptosis after fluorescence staining Apoptotic rate of A549 cells was 58.7% and 42.7%, respectively. MAP30 and MAP30 and MAP30 MAP30-PEG conjugates showed an activation effect on intracellular caspase 3 during apoptosis. Conclusions Both MAP30 and MAP30-PEG conjugates can significantly inhibit the proliferation of A549 cells. Both of them can induce apoptosis through caspase3 pathway.