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目的探讨三氧化二砷(AS_O_3)对人骨肉瘤细胞(HOS,MG63)自噬现象的影响。方法用MTr法观察AS_2O_3对HOS、MG63细胞活力的作用;用透射电镜、免疫荧光染色、Western blot检测HOS、MG63细胞基础自噬以及AS_2O_3诱导细胞自噬的情况。结果透射电镜、免疫荧光染色、Western blot结果显示MG63基础自噬水平明显高于HOS(P<0.01);AS_2O_3抑制MC63细胞活力的IC_(50)值15.42μmol/L明显大于AS_2O_3抑制HOS细胞的活力的IC_(50)值1.067μmol/L,多耐药株MG63较HOS对AS_2O_3耐药;检测LC3-Ⅱ及PARP蛋白的表达水平发现,随着HOS自噬水平的增加,HOS的凋亡逐渐增加,而AS_2O_3通过促进MG63的保护性自噬延缓了凋亡的发生。结论 AS_2O_3可引起骨肉瘤细胞自噬,对不同的骨肉瘤细胞自噬的影响各不相同。对于化疗耐药的骨肉瘤细胞MG63,AS_2O_3诱导的自噬是保护性自噬,自噬延缓了凋亡的发生;而对于化疗敏感的HOS细胞,AS_2O_3诱导的自噬促进了凋亡的发生。
Objective To investigate the effect of arsenic trioxide (AS_O_3) on autophagy in human osteosarcoma cells (HOS, MG63). Methods The effect of AS_2O_3 on the viability of HOS and MG63 cells was observed by MTT assay. The basic autophagy of HOS and MG63 cells and autophagy induced by AS_2O_3 were detected by transmission electron microscopy, immunofluorescence staining and Western blot. Results The results of transmission electron microscopy, immunofluorescence staining and Western blot showed that the basal autophagic level of MG63 was significantly higher than that of HOS (P <0.01). The IC 50 value of AS_2O_3 inhibiting the viability of MC63 cells was significantly higher than that of AS_2O_3 (15.42μmol / L) The IC 50 value was 1.067μmol / L, multi-drug resistant strain MG63 was more resistant to AS_2O_3 than HOS. The expression levels of LC3-Ⅱ and PARP protein were detected. The apoptosis of HOS increased gradually with the increase of HOS autophagy , While AS_2O_3 delayed the apoptosis by promoting the protective autophagy of MG63. Conclusion AS_2O_3 can induce autophagy in osteosarcoma cells and affect the autophagy in different osteosarcoma cells. Chemotherapy-resistant osteosarcoma cells MG63, AS_2O_3-induced autophagy is a protective autophagy, autophagy delayed the occurrence of apoptosis; while for the chemotherapy-sensitive HOS cells, AS_2O_3-induced autophagy promoted apoptosis.