邻-氨基酚去甲表鬼臼毒素抗骨肉瘤SOSP-9607细胞作用及机理研究

Anti-osteosarcoma effects and mechanisms of 4-o-amino-phenol-4′-demethyl epipodophyllotoxin ether on SOSP-9607 cells

目的:研究邻-氨基酚去甲表鬼臼毒素(ODE)抗骨肉瘤SOSP-9607细胞作用及机理.方法:采用MTT比色法测定体外抗骨肉瘤SOSP-9607细胞作用,透射电子显微镜观察细胞凋亡形态学;采用激光共聚焦显微镜测定SOSP-9607细胞内[Ca2+],[Mg2+],pH值和线粒体膜电位(MMP)的荧光强度;免疫细胞化学技术检测细胞内Bc1-2和Bax表达.结果:ODE时间、浓度依赖性地抑制SOSP-9607细胞生长,诱导细胞凋亡具有典型的凋亡形态特征如细胞膜表面微绒毛消失、核固缩、核碎裂等;剂量依赖性地增加SOSP-9607细胞内Ca2+,Mg2+浓度而降低细胞内pH值和线粒体膜电位;使SOSP-9607细胞内Bcl-2蛋白表达下降而Bax蛋白表达增加.结论:邻-氨基酚去甲表鬼臼毒素在体外实验中能明显抑制骨肉瘤SOSP-9607细胞生长,其作用机制可能与改变细胞内环境稳态及Bcl-2和Bax蛋白表达而诱导细胞凋亡有关.

AIM： To investigate the anti-osteosarcoma effects and mechanisms of 4-o-amino-phenol-4＇-demethyl epipodophyllotoxin ether （ODE） on SOSP-9607 cells. METHODS： The antiosteosarcoma effect was determined by microcuhure tetrazolium （MTT） assay; Apoptosis features were observed by transmission electron microscopy. Intracellular Ca^2＋ , Mg^2＋ concentrations, pH value and mltochondrial membrane potential （MMP） were detected by confocal laser scanning microscope. Immunohistochemistry was performed to analyze the changes of Bcl-2 and Bax expression. RESULTS： ODE inhibited the proliferation of SOSP- 9607 cells in a time-and concentration-dependent manner. Typical apoptotic features, such as nuclear fragmentation, chromatin condensation and microvilli disappearance, were observed in SOSP- 9607 cells treated with ODE. Meanwhile, the intracellular Ca^2＋ and Mg^2＋ concentrations were greatly increased whereas pH value and MMP were markedly reduced in SOSP-9607 cells in a concentration-dependent manner after treatment with ODE. The decrease of Bcl-2 and increase of Bax expression were detected in SOSP- 9607 cells after ODE treatment. CONCLUSION： The anti-osteosareoma mechanisms of ODE are attributed to the induction of apoptosis through increasing intracellular Ca^2＋ and Mg^2＋ concentrations, and reducing pH value and MMP, and changing Bcl-2 and Bax expressions.