蛋白酶体抑制剂协同组蛋白去乙酰化酶抑制剂诱导T细胞淋巴瘤凋亡及其与蛋白聚集体关系的研究  

Synergistic Effects of Proteasome Inhibitor and Histone Deacetylase Inhibitor on Apoptosis and Aggresome Formation in T Lymphoma Cells

作　　者：姜晓星[1] 张群岭[1] 陈新宇[2] 邬维礼[1] 沈志祥[1] 赵维莅[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、医学基因组学国家重点实验室,上海200025 [2]上海交通大学医学院细胞生物学教研室,上海200025

出　　处：《中国实验血液学杂志》2009年第5期1215-1219,共5页Journal of Experimental Hematology

基　　金：国家高技术研究发展计划(编号863,2006AA02A301);国家自然科学基金(编号30750335);教育部新世纪优秀人才支持计划;上海市科委国际合作项目(编号08410708800);霍英东青年教师基金(编号111035)

摘　　要：本研究探讨蛋白酶体抑制剂硼替唑米(bortezomib)和组蛋白去乙酰化酶抑制剂辛二酰苯胺异羟肟酸(suberoylanilide hydroxamic acid,SAHA)对T淋巴瘤细胞株Jurkat和Hut78的协同诱导凋亡作用,以及两药联合对蛋白聚集体形成的影响。硼替唑米(10nmol/L)单药或硼替唑米(10nmol/L)联合SAHA(2μmol/L)处理Jurkat和Hut78细胞,应用台盼蓝拒染法计数细胞生长抑制率;细胞涂片观察细胞形态学改变;流式细胞术检测细胞凋亡变化;透射电子显微镜观察细胞凋亡和聚集体形成的超微结构特征。研究结果表明,与对照组和单用硼替唑米组比较,两药合用能够显著抑制Jurkat和Hut78细胞生长,促进细胞凋亡。流式结果显示,两药合用组的Jurkat和Hub78细胞凋亡细胞比例分别为(41.8±4.7)%和(72.7±11.7)%,显著高于对照组[(3.6±1.3)%和(7.0±1.9)%]和单用硼替唑米组[(6.3±2.3)%和(18.7±9.2)%](p均<0.01)。超微结构显示,单用硼替唑米组细胞内多为典型的聚集体结构,两药合用组细胞内聚集体密度降低,数量减少甚至消失,且凋亡细胞明显增多。结论:蛋白酶体抑制剂联合组蛋白去乙酰化酶抑制剂能有效诱导T淋巴瘤细胞凋亡,两药具有协同作用。硼替唑米促进聚集体形成,SAHA能破坏聚集体结构,是增强硼替唑米促凋亡效应的可能机制之一。The aim of the study was to explore the synergistic effect of the proteasome inhibitor bortezomib （bor） and the histone deacetylase inhibitor suberoylanilide hydroxamic acid （SAHA） on apoptosis of T lymphoma cell lines Jurkat and Hut78, and on the formation of aggresome. Jurkat and Hut78 cells were treated with bor （ 10 nmol/L） or bor （ 10 nmol/L） combined with SAHA （2μmol/L） respectively. Cell growth inhibition was estimated by trypan blue dye exclusion test. Cell morphology was evaluated by light microscopy with Wright＇s staining of cytocentrifuge preparations. Cell apoptosis was analyzed by flow cytometry. Ultrastructure of cell apoptosis and aggresome were observed by transmission electron microscopy. The results showed that proliferation of both Jurkat and Hut78 cells was significantly inhibited in the bor ± SAHA group, as compared with the control group and the bor alone group. Flow cytometric analysis confirmed that the percentage of apoptosis in Jurkat and Hut78 cells in the bor ± SAHA group （41.8 ±4.7% and 72.7 ± 11.7% respectively） was remarkably higher than those in the control group （3.6 ± 1.3% and 7.0 ± 1.9% respectively） and the bor alone group （6.3 ± 2.3% and 18.7 ± 9.2% respectively） （p 〈 0.01 ）. Ultrastructure examination revealed that typical aggresomes in cells could be observed in bor alone group. The combination of bor and SAHA diminished both the amount and density of aggresomes, or even eliminated them, accompanied by the increased rate of apoptosis. It is concluded that proteasome inhibitor combined with histone deacetylase inhibitor synergically induces T lymphoma cell apoptosis. Bortezomib stimulates the formation of aggresome, while SAHA destroys this aggresome structure, which may be one of the mechanisms underlying the enhancement of bortezomib-induced apoptosis.

关 键 词：蛋白酶体抑制剂 硼替唑米 组蛋白去乙酰化酶抑制剂 辛二酰苯胺异羟肟酸 T细胞淋巴瘤 细胞凋亡 蛋白聚集体 

分 类 号：R733.1[医药卫生—肿瘤] R979.1[医药卫生—临床医学]