摘要
目的评估衣霉素单独及联合肿瘤坏死因子相关的凋亡诱导配体(TRAIL)对11q23异常白血病细胞株凋亡的影响,并探讨其联合作用后细胞凋亡的通路。方法以t(11;19)急性淋巴细胞白血病细胞株KOCL44细胞株为研究对象,设立实验组及对照组,通过台盼蓝活性测定观察细胞存活率;用FITC标记的An2nexin-V/PI染色,流式细胞仪检测早期凋亡及药物作用后细胞表面死亡受体的表达;Western blotting法检测Bip、CHOP、caspase-3的蛋白表达水平,探讨凋亡通路。结果衣霉素与TRAIL联合处理KOCL44细胞株,对细胞生长均有抑制作用,两药联合有协同作用;Annexin-V/PI检测结果与此一致;衣霉素及TRAIL联合处理KOCL44细胞株后细胞表面DR4未见明显表达,而DR5受体表达明显增强;western blotting检测caspase-3、Bip、CHOP的表达,两药联合作用可见Bip、CHOP表达,并随作用时间延长表达增强。结论衣霉素及TRAIL联合作用KOCL44细胞株后,细胞凋亡明显;衣霉素通过CHOP凋亡通路及Bip介导的UPR上调细胞表面TRAIL死亡受体DR5的表达,以增强11q23异常急性淋巴细胞白血病细胞株对TRAIL诱导凋亡的敏感性。
Objective To assess the effect of tunicamycin alone and with TRAIL on the apotosis of leukemia cell line with 11q23 abnormality,and to explore the possible apoptosis pathway.Methods t（11;19）-ALL cell line KOCL44 was enrolled in the study.1640 culture medium,tunicamycin or TRAIL alone,both tunicamycin and TRAIL were added in blank group,negative control groups and experimental groups,respectively.The cell survival rate was measured by typan blue staining.Early apoptosis was detected with flow cytometry by FITC labeled Annexin-V/PI staining.The cell surface expression of death receptors was detected with flow cytometry.The protein level of Bip,CHOP and caspase-3 was detected by Western blotting.Results Cell growth were significantly inhibited by combination of tunicamycin with TRAIL（P0.05）;Annexin-V/PI staining was observed the same tendency;Cell surface expression of DR5（not DR4） was significantly induced by the combination of tunicamycin and TRAIL;Upon western blotting,Bip and CHOP were activated after treatment with tunicamycin plus TRAIL and the expression was time dependent.Conclusion Tunicamycin enhances the sensitivity of KOCL44 to TRAIL induced apoptosis through Bip mediated UPR,as well as CHOP apoptosis pathway to up-regulating the cell surface expression of death receptor DR5.
出处
《宁夏医科大学学报》
2011年第9期815-818,822,共5页
Journal of Ningxia Medical University
基金
宁夏医科大学科研项目(XT200817)
