肿瘤坏死因子相关凋亡诱导配体在B淋巴瘤细胞株中的敏感性及诱导细胞凋亡机制  

Study on the Sensitivity of TRAIL in B Lymphoma Cell Lines and TRAIL-induced Apoptosis

作　　者：卿晓玲 陈自敏[1] 陈瑜[1] 王洪[1] 李艳[1] 余赟[2] 余小凤[1] 尹春莉 姚双群 田茂岑 杨平[2] 李敏惠[3] Qing Xiaoling Chen Zhimin Chen Yu Wang Hong Li Yan Yu Yun Yu Xiaofeng Yin Chunli Yao Shuangqun Tian Maocheng Yang Ping Li Minhui(School of Biomedical Sciences, Chengdu Medical College, Chengdu 610083, China School of Basic Medical Sciences, Chengdu Medical College, Chengdu 610083, China Center of Scientific Research , Chengdu Medical College, Chengdu 610083, China)

机构地区：[1]成都医学院生物医学系,成都610083 [2]成都医学院基础医学院,成都610083 [3]成都医学院科研实验中心,成都610083

出　　处：《成都医学院学报》2016年第4期413-416,442,共5页Journal of Chengdu Medical College

基　　金：四川省教育厅项目(No:13ZB0220);四川省科技厅项目(No:2015JY0205);四川省卫生厅项目(No:130302;130298);国家级大学生创新项目(No:201313705007;201313705003;201413705005);成都医学院科研基金(No:CYZ11-005)

摘　　要：目的探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)在B淋巴瘤细胞株中的敏感性及TRAIL通过线粒体信号通路诱导人B淋巴瘤细胞株Ramos凋亡的机制。方法采用CCK-8法检测肿瘤细胞增殖,流式细胞术分析细胞线粒体膜电位、细胞表面受体DR4/DR5表达及细胞凋亡率变化,并用Western Blot测定Ramos细胞经TRAIL作用24h后相关凋亡蛋白的表达水平。结果供试的4株B淋巴瘤细胞中,Ramos细胞为TRAIL敏感株,Ramos细胞表面受体DR4/DR5表达水平明显高于TRAIL耐药株;TRAIL抑制Ramos细胞增殖通过诱导细胞凋亡介导;细胞调亡伴随线粒体膜电位明显变化;Western Blot免疫印迹结果显示,药物作用后,细胞内相关凋亡信号分子包括Caspase-9,Caspase-3,PARP蛋白等均明显活化。结论 4株B淋巴瘤细胞株对TRAIL敏感性不同,并与DR4/DR5表达相关;TRAIL抑制Ramos细胞增殖,诱导细胞凋亡,引起线粒体膜电位明显下降,活化Caspase-9及下游凋亡蛋白;线粒体调亡途径是介导TRAIL诱导Ramos细胞凋亡的途径之一。Objective To investigate the sensitivity of TRAIL in B lymphoma cell lines and the mechanism of Ramos apoptosis induced by TRAIL via the mitochondrial signaling pathways.Methods The CCK-8assay was adopted to detect the tumor cell proliferation.The flow cytometry was used to analyze the cell mitochondrial membrane potential,the cell surface receptors DR4/DR5 expressions and the changes of apoptosis.The Western Blot Test was employed to test the expression levels of apoptosis protein after 24 hours with TRAIL stimulation.Results Among the 4Blymphoma cell lines,Romas cells are the sensitive cell strain to TRAIL and the DR4/DR5 expression levels were significantly higher than the drug-resistant strains of TRAIL.TRAIL inhibited Ramos cell proliferation by inducing the cell apoptosis accompanied by the significant changes of mitochondrial membrane potential.The results of Western-Blot Analysis indicated that the apoptotic signal molecules such as Caspase-9,Caspase-3and PARP were significantly activated.Conclusion The sensitivity of the 4experimental B lymphoma cell lines to TRAIL is different and it is related to the DR4/DR5 expressions.TRAIL inhibits cell proliferation of Ramos Cells,induces their apoptosis,decreases the mitochondrial membrane potential significantly,and activates Caspase-9and Caspase-3.Mitochondrial apoptosis pathway is one of the ways to mediate the apoptosis of Ramos cells induced by TRAIL.

关 键 词：TRAIL B淋巴瘤 细胞凋亡 线粒体途径 

分 类 号：R730.5[医药卫生—肿瘤]