羟基磷灰石纳米颗粒对骨肉瘤MG63抑制作用的体外实验研究

Experimental Study of the Inhibitory Effect of Hydroxyapatite Nanoparticles on Osteosarcoma MG63 in vitro

目的探讨羟基磷灰石(Hydroxyapatite,HAP)纳米颗粒对骨肉瘤MG63的作用及相关机制。方法 MTT法观察不同浓度HAP纳米颗粒对MG63的抑制作用,测定HAP纳米粒子对MG63的时效曲线,丫啶橙/溴化已啶细胞荧光染色及流式细胞术检测HAP纳米颗粒对MG63细胞凋亡的影响。结果 MTT法检测发现HAP纳米颗粒浓度在10-400 mg/L范围内,随着浓度的增高HAP纳米颗粒对MG63的抑制作用在72 h内逐渐增强,以后逐渐达到平台期。形态学观察和流式细胞术检测可见HAP纳米粒子有诱导细胞凋亡的趋势,实验组和对照组的细胞凋亡率(%)分别为19.03±5.80和12.32±3.84,两组间差别显著(P<0.05);实验组G0/G1期细胞明显多于对照组,而S期细胞明显少于对照组,两组间差别显著(P<0.05)。结论 HAP纳米颗粒可以诱导肿瘤细胞凋亡,对骨肉瘤MG63有明显的抑制作用。

Objective To investigate the effect of hydroxyapatite（HAP） nanoparticles on the osteosarcoma cell line MG63,and the related mechanism was discussed initially.Methods The inhibition of different concentraten HAP nanoparticles on MG63 was observed by MTT assay,and the time-activity curve and dose-effect curve were measured.The apoptosis of MG63 was observed by the fluorescence dyed with Acridine orange and Ethidium bromide,and the rate of apoptosis was also detected by flow cytometry.Results The inhibition rate of MG63 was gradually increased with the variable concentraten of HAP nanoparticles within 72 hours.The morphological detections demonstrated that the HAP nanoparticles could induce the apoptosis of MG63 cell.The apoptosis rate of HAP nanoparticles group and control group was 19.03±5.80 and 12.32±3.84,respectively.There was significant difference between two groups（P〈0.05）.In HAP nanoparticles group,the amount of cells at G0/G1 phase were superior than control group,and the amount of cells at S phase were inferior than control group（P〈0.05）.Conclusion The HAP nanoparticles could induce the apoptosis of the carcinoma cells,and inhibit the growth of MG63.