Velcade诱导多发性骨髓瘤细胞株U266细胞凋亡研究

为了研究velcade诱导多发性骨髓瘤细胞株U266细胞凋亡效应及机制，采用台盼蓝拒染法检测2、10、50和100nmol／L velcade处理U266细胞活率，用流式细胞术分析Annexin—V、线粒体跨膜电位（△ψm）和氧自由基（ROS），用半定量RT—PCR法检测bcl-2 mRNA的表达。结果表明：velcade抑制U266细胞增殖；诱导U266细胞凋亡，24小时Annexin—V阳性细胞比例增高，△ψm降低；12小时时活性氧明显增高，荧光强度增强；抗凋亡基因bcl-2表达降低。结论：velcade对U266细胞具有增殖抑制和杀伤作用。其可通过内源性细胞凋亡信号通路诱导U266细胞凋亡。

Multiple Myeloma Cell Line U266 Apoptosis Induced by Velcade

To investigate the effect of velcade on multiple myeloma cell line U266 apoptosis and its mechanism, cell viability was estimated by trypan blue dye exclusion. Annexin-V, mitochondrial transmembrane potential (delta psi m) and reactive oxygen species (ROS) labeled by DCFHDA were examined by flow cytometry, the expression of bcl-2 mRNA was detected by semi-quantitative RT-PCR. The results showed that the velcade inhibited the growth of U266 cells and reduced cell viability accompanied by appearance of morphologic characteristics of apoptosis. Velcade at 50 nmol/L increased Annexin V positivity and fluorescence intensity of DCF because of ROS generation while it decreased the delta psi m of U266 cells. Expression of anti-apoptotic gene bcl-2 mRNA also decreased. It is concluded that velcade inhibited the growth and reduce cell viability of U266 cells. Velcade can induce U266 cells apoptosis by intrinsic cell apoptotic pathway.