CAJ | 학술논문

目的：探讨三氧化二砷(AS2O3)对人骨肉瘤细胞(HOS，MG63)自噬现象的影响。方法用MTT法观察AS2O3对HOS、MG63细胞活力的作用；用透射电镜、免疫荧光染色、Westernblot检测HOS、MG63细胞基础自噬以及AS2O3诱导细胞自噬的情况。结果透射电镜、免疫荧光染色、Westernblot结果显示MG63基础自噬水平明显高于HOS(P＜0.01)；AS2O3抑制MG63细胞活力的IC50值15.42μmol/L明显大于AS2O3抑制HOS细胞的活力的IC50值1.067μmol/L，多耐药株MG63较HOS对AS2O3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AS2O3通过促进MG63的保护性自噬延缓了凋亡的发生。结论 AS2O3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，AS2O3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，AS2O3诱导的自噬促进了凋亡的发生。
목적：탐토삼양화이신(AS2O3)대인골육류세포(HOS，MG63)자서현상적영향。방법용MTT법관찰AS2O3대HOS、MG63세포활력적작용；용투사전경、면역형광염색、Westernblot검측HOS、MG63세포기출자서이급AS2O3유도세포자서적정황。결과투사전경、면역형광염색、Westernblot결과현시MG63기출자서수평명현고우HOS(P＜0.01)；AS2O3억제MG63세포활력적IC50치15.42μmol/L명현대우AS2O3억제HOS세포적활력적IC50치1.067μmol/L，다내약주MG63교HOS대AS2O3내약；검측LC3-II급PARP단백적표체수평발현，수착HOS자서수평적증가，HOS적조망축점증가，이AS2O3통과촉진MG63적보호성자서연완료조망적발생。결론 AS2O3가인기골육류세포자서，대불동적골육류세포자서적영향각불상동。대우화료내약적골육류세포MG63，AS2O3유도적자서시보호성자서，자서연완료조망적발생；이대우화료민감적HOS세포，AS2O3유도적자서촉진료조망적발생。
Objective To explore the effects of arsenic trioxide ( AS2O3 ) on autophagy in human osteosarcoma cells ( HOS, MG63 ). Methods The effects of AS2O3 on the viability of HOS and MG63 cells were detected by 3-( 4,5-Dimethylthiazol-2-yl )-2,5-diphenyltetrazolium bromide ( MTT ) assay. The basal and AS2O3-induced levels of autophagy were observed by transmission electron microscope, immunofluorescent staining and Western blot assay. Results The examination results of transmission electron microscope, immunolfuorescent staining and Western blot assay showed the basal level of autophagy of MG63 cells was higher than that of HOS cells ( P<0.01 ). The half maximal inhibitory concentration ( IC50 ) value of AS2O3 on MG63 cells was 15.42 μmol/L, which was obviously higher than 1.067μmol/L of IC50 value of AS2O3 on HOS cells. HOS cells were more sensitive to AS2O3 than MG63 cells. The protein expression levels of microtubule-associated protein 1 light chain 3 ( LC3 )-II and poly-ADP-ribose polymerase ( PARP ) were detected. The apoptosis was gradually serious with the improvement of AS2O3-induced autophagy in HOS cells, but AS2O3-induced autophagy protected the MG63 cells from apoptosis. Conclusions The AS2O3-induced autophagy plays different roles in osteosarcoma cells, and the apoptosis of tumor cells can be promoted so as to achieve the aim of erase osteosarcoma. For MG63 cells of chemotherapy resistance, the AS2O3-induced autophagy is protective, and the apoptosis will be delayed by autophagy. As to HOS cells of chemotherapy sensitivity, the apoptosis will be induced by autophagy.