青蒿素衍生物SM1044诱导弥漫大B细胞淋巴瘤细胞株SU-DHL-4凋亡的机制研究  

The mechanism of induction of apoptosis of diffuse large B-cell lymphoma cell line SU-DHL-4 byartemisinin derivatives SM1044

作　　者：宋志群[1] 赵玲玲[1] 崔雯[1] 苗圣超 王瑾[1] 李英[2] 糜坚青[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院血液科上海血液学研究所,上海200025 [2]中国科学院上海药物研究所,上海201203

出　　处：《内科理论与实践》2014年第4期274-279,共6页Journal of Internal Medicine Concepts & Practice

基　　金：国家重点基础研究发展计划(973计划)(项目编号:2010CB529203);上海市教育委员会科研创新重点项目(项目编号:13zz088)

摘　　要：目的:研究青蒿素衍生物SM1044诱导弥漫大B细胞淋巴瘤细胞株SU-DHL-4凋亡的相关机制。方法:流式细胞术检测SU-DHL-4细胞的凋亡情况;蛋白质印迹(Western blotting)检测凋亡相关蛋白和内质网应激相关蛋白的表达;实时PCR检测内质网应激相关基因的表达;免疫荧光技术检测细胞内钙离子的荧光强度。结果:SM1044可诱导SU-DHL-4细胞凋亡,且具有剂量依赖性(r=0.98,P=0.02);促进胱天蛋白酶3剪切片段及多腺苷二磷酸核糖聚合酶剪切片段的产生,差异有统计学意义(P<0.05);SM1044可使SU-DHL-4细胞中内质网应激相关基因和蛋白的表达显著上调,差异有统计学意义(P<0.05);SM1044可促进细胞内钙离子的水平显著升高。钙离子螯合剂1,2-二(2-氨基苯氧基)乙烷-N,N,N’,N’-四乙酸四乙酰氧甲基酯(BAPTA-AM)和SM1044联用组,抗CCAAT增强子结合蛋白(C/EBP)环腺苷酸反应元件结合转录因子同源蛋白(CHOP)基因的表达改变是对照组的(2.494±0.289)倍,明显低于SM1044单用组的(4.204±0.429)倍,差异有统计学意义(P<0.01);CHOP的表达也由SM1044单用组(5 734.46±713.66)下调为联用组的(2 006.17±710.43),差异有统计学意义(P<0.01);同时钙离子螯合剂BAPTA-AM也可显著下调SM1044诱导的胱天蛋白酶3剪切片段及多腺苷二磷酸核糖聚合酶剪切片段的表达,差异有统计学意义(P<0.001)。结论:SM1044可以诱导SU-DHL-4细胞发生凋亡,其机制可能与SM1044促进细胞内钙离子水平升高和内质网应激相关。Objective To investigate the mechanism of induction of apoptosis of diffuse large B-cell lymphoma cell line SU-DHL-4 by artemisinin derivatives SM1044. Methods Flow cytometry was used to examine cell apoptosis. The expression of apoptosis and endoplasmic reticulum stress related proteins was detected by Western blotting. The transcriptional level of endoplasmie retieulum stress related genes was determined with quantitative real-time PCR. Immunofluorescenee was performed to measure the level of intracellular calcium. Results SM1044 could induce apoptosis of SU-DHL-4 in a dose dependent manner （r=0.98, P=0.02）, and the expression of apoptosis associated proteins, such as cleaved-caspase 3 and eleaved-poly adenosine diphosphate （ADP）-ribose polymerase （PARP） was increased, and the difference was statistically significant （P〈0.05）. The expression of endoplasmic reticulum stress related genes and proteins was significantly enhanced by SM1044 （P〈0.05）, and so was the level of intracellular calcium. In calcium ion chelator 1, 2-his （2-aminophenoxy）ethane-N, N, N＇, N＇tetraacetic acid tetraaeetoxymethyl ester（BAPTA-AM） combined with SM1044 group, the change of expression of CCAAT/enhancer-binding protein homologous protein（CHOP） gene was （2.494±0.289）folds of that of control group, which was significantly lower than that of SM10d4 alone group （4.204±0.429） folds, and the difference was statistically significant （P〈0.01）. The expression of CHOP was （5 734.46±713.66） in SM1044 alone group, and was decreased to （2 006.17±710.43） when combined with BAPTA-AM, the difference was statistically significant （P〈 0.01）. Calcium ion chelator BAPTA-AM could inhibit the expression of apoptosis associated proteins, such as the cleavedcaspase 3 and cleaved-PARP induced by SM1044, the difference was statistically significant （P〈0.001）. Conclusions SM1044 could significantly induce the apoptosis of SU-DHL-4, which might be related to the elevated level of intrac

关 键 词：青蒿素衍生物 弥漫大B细胞淋巴瘤 SU-DHL-4细胞株 细胞凋亡 

分 类 号：R733.1[医药卫生—肿瘤]