羟基磷灰石纳米粒子诱导人肝癌BEL-7402细胞凋亡的机制  

Mechanism of hydroxyapatite nanoparticles inducing apoptosis of human hepatoma cell line BEL- 7402

作　　者：唐胜利[1] 刘志苏[1] 钱群[1] 江从庆[1] 艾中立[1] 

机构地区：[1]武汉大学中南医院普外科,430071

出　　处：《中华实验外科杂志》2012年第4期660-662,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30471689）

摘　　要：目的探讨羟基磷灰石纳米粒子（nano—HAP）诱导肝癌细胞凋亡的机制。方法羟基磷灰石纳米粒子处理BEL-T402细胞后，免疫印迹法检测p53、c—myc蛋白表达水平；比色法检测半胱氨酸天冬氨酸蛋白酶-3（Caspase-3）活性；端粒重复序列扩增-酶链免疫吸附反应法（TRAP-ELISA）方法测定细胞内端粒酶活性的变化。结果羟基磷灰石纳米粒子处理BEL-7402细胞使抑癌基因p53的表达上升、癌基因c-myc的表达下降。随羟基磷灰石纳米粒子浓度的递增（0、50、75、100、150、200mg／L），端粒酶活性降低（1．94、1．26、1．21、1．14、0．86、0．59），随着作用时间延长（0、12、24、36、48h）端粒酶活性降低（1．94、1．73、1．57、1．43、1．26），并激活Caspase-3。结论羟基磷灰石纳米粒子能通过上调p53、下调c-myc基因表达，抑制端粒酶活性及激活Caspase-3等途径诱导肝癌细胞凋亡。Objective To investigate the mechanism of apoptosis induction bu nano hydroxyapatite nano-particles （HAP）. Methods Western blotting was used to detect the expression of anti-oncogene p53 and that of oncogene c-myc. The catalytic activity of the Caspases was measured by using a colofimetric as- say. Telomeric repeat amplification protocol （TRAP） combined with enzyme-linked immunosorbent assay （ELISA） was used to detect the inhibitory rate of telomerase activity. Results The treatment of BEL-T402 cells with nano-HAP decreased the expression of p53 protein, but increased the expression of c-myc pro- tein. The Caspase-3 activity was increased during nano-HAP-induced apoptosis, while the telomerase activ- ity was decreased by nano-HAP in a dose- and time-dependent manner. With the progressive increases in concentrations of nano-HAP （0, 50, 75, 100, 150, 200 mg/L）, the telomerase activity was down-regula- ted （ 1.94, 1.26, 1.21, 1.14, 0. 86, 0. 59, respectively, all P 〈 0. 01 ）. At the concentration of 50 mg/ L nano-HAP at different time points （0, 12, 24, 36, 48 h）, the telomerase activity was down-regulated （ 1.94, 1.73, 1.57, 1.43, 1.26, respectively, all P 〈 0.01 ）. Conclusion These results suggested that nano-HAP-induced apoptosis was mediated by altering the expression of anti-oncogene p53 and oncogene c-myc, activating Caspase-3 and inhibiting activity of telomerase.

关 键 词：羟基磷灰石纳米粒子 癌 肝细胞 P53 半胱氨酸天冬氨酸蛋白酶-3 端粒酶 

分 类 号：R735.7[医药卫生—肿瘤]