冬凌草甲素诱导人多发性骨髓瘤LP-1细胞凋亡的实验研究  

Experimental Research on the Mechanisms of Human Multiple Myeloma LP-1 Cell Apoptosis Induced by Oridonin

作　　者：赵晶[1] 张梅[1] 陈颖[1] 

机构地区：[1]西安交通大学医学院第一附属医院血液内科,西安710061

出　　处：《中国中西医结合杂志》2012年第12期1642-1646,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金青年基金资助项目(No.81000218)

摘　　要：目的探讨冬凌草甲素(Oridonin)对LP-1细胞的诱导凋亡作用及其机制。方法采用不同浓度冬凌草甲素处理人多发性骨髓瘤(multiple myeloma,MM)LP-1细胞,通过MTT比色法检测LP-1细胞的增殖活性,Annexin V/PI双染色法检测LP-1细胞的凋亡效应,透射电子显微镜观察药物作用后细胞超微结构的变化,RT-PCR检测凋亡相关基因mRNA表达变化。结果冬凌草甲素抑制LP-1细胞增殖呈时间和剂量依赖性;Annexin V/PI双染色法检测显示,随着作用药物浓度的增加以及药物作用时间的延长,LP-1细胞凋亡率显著增加。通过透射电子显微镜可见药物作用后的细胞呈现核染色质边集,线粒体肿胀等细胞凋亡的表现。冬凌草甲素作用后LP-1细胞中PDCD5、Bid mRNA表达上调,Bcl-2、NF-κB mRNA表达下调。结论冬凌草甲素通过上调Bid及下调Bcl-2 mRNA表达降低线粒体膜电位、触发LP-1细胞的线粒体凋亡途径诱导细胞凋亡;亦可作为NF-κB活性抑制剂阻断NF-κB激活,诱导细胞凋亡、抑制细胞增殖。其中PDCD5作为凋亡促进剂的作用仍需进一步的深入研究。Objective To explore the effects of oridonin （Ori） on human multiple myeloma （MM） cell line LP-1 apoptosis and its mechanisms. Methods The human MM LP-1 cells were incubated in vitro by different concentrations of Od. The proliferation activities of LP-1 cells were detected using MT＇r assay. The apoptosis rate of LP-1 cells was detected using Annexin V/PI double staining method. The ultrastructural changes of LP-1 cells were observed under transmission electron microscope （TEM） after they were treated with Ori. The mRNA ex- pression of apoptosis correlated genes were detected using Real-time PCR. Results Ori inhibited the proliferation of LP-1 cells in a dose- and time-dependent way. Results of Annexin V/PI double staining method showed that, along with increased drug concentration and prolonged drug action time, the apoptosis rate of LP-1 cells signifi- cantly increased. Under TEM, chromatin margination and mitochondrial swelling could be seen in LP-1 cells after they were treated by Ori. The mRNA expressions of PDCD5 and Bid were up-regulated, and those of Bcl-2 and NK-KB were down-regulated after action of Ori. Conclusions Ori induced cell apoptosis by up-regulating the mR- NA expression of Bid and down-regulating the mRNA expression of Bcl-2 to decrease the mitochondrial mem- brane potential, trigger mitochondrial apoptosis way of LP-1 cells. Ori, also as the inhibitor of NF-KB activities, blocked the NF-KB activation, induced cell apoptosis, and inhibited the cell proliferation. Of them, it is necessary to further study the role of PDCD5 as an apoptosis promoter.

关 键 词：冬凌草甲素 多发性骨髓瘤 LP-1细胞 细胞凋亡 

分 类 号：R733.3[医药卫生—肿瘤]