马钱子碱经死亡受体途径诱导人类多发性骨髓瘤U266细胞凋亡  

Apoptosis of U266 cells induced by brucine through death-receptor pathway

作　　者：李志华[1] 马艳萍[1] 王艺华[1] 冯静[1] 

机构地区：[1]山西医科大学第二医院血液科,太原030001

出　　处：《白血病．淋巴瘤》2010年第12期724-727,731,共5页Journal of Leukemia & Lymphoma

摘　　要：目的 探讨马钱子碱诱导人类多发性骨髓瘤U266细胞凋亡的机制.方法 通过MTT法、流式细胞术、RT-PCR法检测及形态学观察马钱子碱诱导人类多发性骨髓瘤U266细胞株凋亡的途径.结果 马钱子碱诱导U266的凋亡呈时间和浓度依赖性,48 h IC50 0.16 mg/ml.Hoechst 33258染色后在荧光显微镜下观察到明显的凋亡小体.Rhodamine 123染色后流式细胞术检测马钱子碱不同浓度下线粒体膜电位的变化差异无统计学意义（P＞0.05）.RT-PCR检测到马钱子碱作用12、24、48 h后Caspase-3表达量增加,其灰度值（0.2597±0.020）、（0.5488±0.016）、（0.6205±0.006）、（0.6533±0.009）（P＞0.05）;分别加入Caspase-8与Caspase-9的特异抑制剂z-IETD-fmk、z-LEHD-fmk后检测Caspase-3的表达变化,其灰度值分别为（0.7118±0.006）、（0.2637±0.003）（P＜0.01）;（0.7182±0.004）、（0.7195±0.003）（P=0.836）,提示Caspase-8被激活.结论 在0.4 mg/ml浓度范围内,马钱子碱具有诱导U266细胞凋亡作用,激活Caspase-8经死亡受体途径诱导U266细胞株凋亡.Objective To investigate the mechanism of the apoptotic effect of brucine on human multiple myeloma. Methods MTT, morphology, flow cytometry, and RT-PCR were used to observe the apoptotic pathways of brucine on human multiple myeloma cell line-U266. Results The apoptotic effect of brucine showed a dose and time dependent manner, 48 hour IC50 0.16 mg/ml. The cells treated with brucine showed significant feature associated with apoptosis by Hoechst 33258 at fluorescence microscope.Mitochondrial membrance potential showed no statistical significant difference in different concentration with Rhodamine 123 by flow cytometry （P 〉0.05）. The Caspase-3 expression detected by RT-PCR was increased at 12, 24 and 48 hours treated with brucine, and its gray value was （0.2597±-0.020）, （0.5488±0.016）, （0.6205±0.006）, （0.6533±0.009） （P 〉 0.05）, detection of added z-IETD-fmk, z-LEHD-fmk （Caspase-8 and Caspase-9specific inhibitor） the expression of Caspase-3, the gray scale values were （0.7118±0.006）, （0.2637±0.003）（P 〈0.01）; and （0.7182±0.004） （0.7195±0.003） （P=0.836）. Caspase-8 was activated. Conclusion Within the 0.4 mg/ml concentration brucine can induce apoptosis in U266 cells. Brucine induced apoptosis on cell line U266 through Caspase-8 activation by death-receptor pathway.

关 键 词：马钱子碱 U266细胞株 细胞凋亡 

分 类 号：R733[医药卫生—肿瘤]