硼替佐米调控内质网应激诱导人急性T淋巴细胞白血病细胞株凋亡的机制探讨

The mechanism of Bortezomib on apoptosis of human T cell acute lymphoblastic leukemia by modulation of endoplasmic reticulum stress

目的:探讨硼替佐米对人急性T淋巴细胞白血病(T cell acute lymphoblastic leukemia,T-ALL)细胞株Molt-4细胞凋亡的影响。方法:硼替佐米(0、100、200和400 nmol/L)处理人急性T淋巴细胞白血病细胞株Molt-4细胞后,运用MTT法测定Molt-4细胞活力;使用Hoechst 33258染色法观察凋亡细胞形态;采用荧光定量PCR法测定Bax以及Bip mRNA水平;运用Western blot法测定Bax以及Bip蛋白水平。结果:100、200和400 nmol/L的硼替佐米处理Molt-4细胞24 h后,可浓度依赖性地降低细胞活力。100、200和400 nmol/L的硼替佐米作用细胞24 h后,Molt-4细胞核发生固缩或裂解。硼替佐米(100、200和400 nmol/L)处理细胞24 h后,Molt-4细胞的Bax mRNA和蛋白表达水平明显增加且可显著上调Bip mRNA和蛋白表达水平。结论:硼替佐米可诱导人急性T淋巴细胞白血病细胞株Molt-4细胞凋亡,作用机制可能与它调控内质网应激有关。

Objective:To investigate the role of Bortezomib on cell apoptosis of human T cell acute lymphoblastic leukemia cell line Molt-4 cells.Methods:After adminstration of Molt-4 cells with different concentrations of Bortezomib(100,200 and 400 nmol/L)for 24 h,cell viability was detected using MTT assay.The morphological of apoptotic cells was observed under a fluorescence microscope using Hoechst 33258 staining.The expression levels of Bax as well as Bip mRNA were measured by quantitative real-time PCR.The expression levels of Bax as well as Bip protein were measured by Western blot.Results:Compared with the 0 nmol/L group,Bortezomib obviously decreased the viability of Molt-4 cells in a dose dependent way.Additionally,after treating with Bortezomib(100,200 and 400 nmol/L),the occurrence of apoptotic morphology and the expression levels of Bax mRNA as well as protein in Molt-4 cells were significantly elevated.The expression levels of Bip mRNA as well as protein in Molt-4 cells were obviously increased after treating by Bortezomib for 24 h.Conclusion:Bortezomib can induce cell apoptosis of human T cell acute lymphoblastic leukemia cell line Molt-4 cells,which may be associated with regulating endoplasmic reticulum stress.