Objective To explore the relationship between autophagy and apoptosis induced by cisplatin(DDP). Methods The effects of autophagy in osteosarcoma were investigated by immunofluorescence staining and electron microscope. Through down-regulation of autophagy with 3-methyladenine(3MA) ,the proliferation inhibition ratio of MG63 cells with DDP was measured by using MTT assay. The expression of autophagy was examined by MDC staining and flow cytometry. The apoptosis of MG63 cells was detected by flow cytometry. Results LC3- Ⅱ green fluorescence diffusely distributed near the cell membrane after treatment with DDP and the autophagosome could be found under the electron microscopy. DDP could induce autophagy and apoptosis. 3MA could suppress autophagy, but had no significant effects on apoptosis. Compared to DDP treatment alone, the apoptosis rate and proliferation ratio of MG63 cells were increased by 3MA pretreatment. 3MA administration following DDP could reduce apoptosis rate and proliferation inhibition ratio in comparison to DDP treatment alone. Conclusion DDP can induce autophagy in the MG63 cells. At the early stage of DDP treatment,autophagy can postpone apoptosis to protect cells. At the late stage of DDP treatment,autophagy can promote cell death as another kind of death with apoptosis together.%目的 研究顺铂(DDP)诱导的骨肉瘤细胞自噬性活化,并探讨自噬与凋亡的关系.方法 利用免疫荧光染色和电镜观察DDP对骨肉瘤细胞自噬的影响;应用3-甲基腺嘌呤(3MA)下调自噬,采用MTT比色法,测定DDP对骨肉瘤细胞增殖抑制率的影响;MDC荧光染色和流式细胞术定量检测骨肉瘤细胞自噬的表达;应用流式细胞仪检测骨肉瘤细胞的凋亡情况.结果 DDP给药后可见LC3-Ⅱ绿色荧光呈点状散在分布于细胞膜附近.电镜下观察到DDP给药后骨肉瘤细胞内自噬体的形成.DDP能诱导自噬和凋亡的发生.3MA能抑制自噬的发生,对凋亡无明显影响.与单独DDP作用相比,3MA提前给药细胞的凋亡率增加,其细胞增殖抑制率也增高;3MA迟后给药细胞的凋亡率略有下降,其细胞增殖抑制率也下降.结论 DDP能诱导MG63细胞发生自噬性活化.DDP作用早期,自噬能延缓凋亡的发生,发挥保护细胞的作用;在DDP作用后期,自噬则作为另一种死亡形式出现,和凋亡一起促进细胞死亡.
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