二膦酸盐对人骨肉瘤细胞诱导凋亡作用的实验研究

目的：探讨二膦酸盐对人骨肉瘤细胞的诱导凋亡作用及其可能的作用机制。方法将人骨肉瘤M G‐63细胞株培养、传代后将细胞分为两组：二膦酸盐400μg／m L干预组、生理盐水空白对照组，孵育72 h后，对两组人骨肉瘤M G‐63细胞株行免疫荧光检测，观察两组细胞中凋亡因子Caspase‐3、Fas的表达；行流式细胞术检测，观察两组细胞的细胞凋亡率。结果二膦酸盐作用于人骨肉瘤MG‐63细胞株72 h后，免疫荧光检测结果可见二膦酸盐400μg／mL干预组细胞中凋亡因子Caspase‐3、Fas大量表达，而生理盐水空白对照组几乎见不到凋亡因子Caspase‐3、Fas的表达；流式细胞术检测结果观察到二膦酸盐400μg／mL干预组细胞的细胞凋亡率为54．00％，远大于生理盐水空白对照组的细胞凋亡率3．10％，差异有统计学意义（ P＜0．05），存在明显的诱导凋亡现象。结论二膦酸盐对体外人骨肉瘤M G‐63细胞株存在很强的诱导凋亡作用。

Apoptotic effects of bisphosphonate inosteosarcoma MG-63 cells in vitro

Objective To observe the effects of bisphosphonate on the inhibit proliferation and the apoptosis effect in osteo‐sarcoma MG‐63 cells in vitro ,explore the phosphonic acid salt of bone sarcoma cells ,induce apoptosis and its possible mechanism . Methods Sixty three osteosarcoma MG‐63 cells were cultured in vitro .After treated with bisphosphonate 400 μg/mL ,without bi‐sphosphonate but normal saline ,they were incubated 72 h after the application of the two group cell immunofluorescence test ;then observe the expression of apoptosis factors Caspase 3 and Fas ;Flow cytometry detection line was used to detect the osteosarcoma cell line MG‐63 cells apoptosis rate of each group .Results 72 h after treatment with bisphosphonate ,the expression of apoptosis factor of Caspase‐3 and Fas in osteosarcoma MG‐63 cells were strongly expressed ,and it was observed by immunofluorescent assay , while in blank control group ,we could barely see the expression of apoptosis factors Caspase‐3 and Fas ;Flow cytometry test results showed that two phosphonic acid salt 400 μg/mL intervention group cell apoptosis rate was 54 .00% ,far more than normal saline blank control group ,of which the apoptosis rate was 3 .10% ,the difference was statistically significant (P<0 .05) ,there is an obvi‐ous phenomenon of induced apoptosis .Conclusion Bisphosphonate has a strong apoptotic effects of bisphosphonate in osteosarcoma MG‐63 cells in vitro .Bisphosphonate can inhibit osteolysis of osteosarcoma MG‐63 cells via regulating the expression of Caspase‐3 , Fas in osteosarcoma MG‐63 cells .Bisphosphonate may serve as a potential therapeutic agent for treatment of osteosarcoma .