硼替佐米联用组蛋白去乙酰化酶抑制剂诱导血液肿瘤细胞凋亡的效率  

Bortezomib in Combination with Histone Deacetylase Inhibitor-induced Apoptosis of Malignant Hematologic Cells

作　　者：孙立石[1] 洪振亚[1] 韩志强[1] 肖敏[1] 曹阳[1] 黄亮[1] 周剑峰[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院血液科,武汉430030

出　　处：《华中科技大学学报（医学版）》2008年第2期185-188,203,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30770914)

摘　　要：目的探讨组蛋白去乙酰化酶（histone deacetylase，HDAC）抑制剂曲古菌素A（trichostatin A，TSA）、Apicidin与Bortezomib单药及联合应用对恶性血液病肿瘤细胞凋亡的影响。方法应用200～2000nmol／L TSA、Apicidin，5～80μmol／L Bortezomib分别处理恶性血液病肿瘤细胞株K562、SHI-1、Jurkat 24h，用流式细胞仪检测细胞凋亡。根据流式细胞术结果选取TSA 800nmol／L，Apicidin 400nmol／L分别与5～80μmol／L Bortezomib联合作用于K562细胞，TSA 1000nmol／L，Apicidin 400nmol／L分别与Bortezomib联合作用于SHI-1细胞，TSA 600nmol／L，Apicidin 600nmol／l，分别与Bortezomib联合作用于Jurkat细胞。以上药物作用24h后应用流式细胞仪检测细胞凋亡并绘制浓度-凋亡曲线。分别以Apicidin 200nmol／L，Bortezomib 20μmol／L及Apicidin 200nmol／L与Bortezomib 5μmol／L联用作用于K562细胞24h，用Western blot检测Bcl-XL蛋白的表达差异。结果TSA、Apicidin、Bortezomib均可促进K562、Jurkat、SHI-1细胞凋亡。TSA、Apicidin分别与Bortezomib联用，可导致K562、SHI-1细胞系凋亡率的明显提高，而对Jurkat细胞系没有明显影响。Bcl-XL蛋白在Apicidin与Bortezomib联用24h组较对照组及单一药物处理组表达水平明显下调。结论小剂量Bortezomib联合TSA或Apicidin应用于K562、SHI-1细胞系，可以明显提高细胞凋亡率，此时，TSA、Apicidin用量较用单药时显著减少。Objective To explore whether the combination of histone deacetylase inhibitor （HDACI） （TSA, Apicidin） and Bortezomib on malignant hematologic cells can bring about stronger influence on apoptosis than single-drug treatment. Methods K562, SHI-1 and Jurkat cells were treated with 200-200 0 nmol/L TSA, Apicidin and 5-80 μmol/L Bortezomib separately and the apoptosis rate was measured by flow cytometry after 24 h. According to the results of single-drug apoptosis rate we acquired, a concentration at 800 nmol/L of TSA, 400 nmol/L of Apicidin was separately combined with Bortezomib to treat K562 cells, a concentration at 1000 nmol/L of TSA, 400 nmol/L of Apicidin with Bortezomib on SHI-1 cells and 600 nmol/L TSA, 600 nmol/L Apicidin with Bortezomib on Jurkat cells and then concentration-apoptosis rate dependent curves were made according to the results. After that, Western blot was employed to detect the expression level of Bcl-XL in proteins extracted from K562 cells treated with 200 nmol/L Apicidin, 20 μmol/L Bortezomib and the combination of 200 nmol/L Apicidin and 5 μmol/ L Bortezomib after 24 h respectively. Results TSA, Apicidin and Bortezomib could induce apoptosis on K562, Jurkat and SHI-1 cells. Besides, Bortezomib was synergistic with TSA or Apicidin greatly on K562 as well as SHI-1 cells and there was no significant effect on Jurkat cells in the combination treatment. The expression level of Bcl-XL was obviously down-regulated in the combination treatment of Apicidin and Bortezomib. Conclusion Small dose of Bortezomib combined with TSA or Apicidin on K562 and SHI-1 cells can greatly enhance apoptosis rate compared with single drug treatment.

关 键 词：组蛋白去乙酰化酶抑制剂 曲古菌素A APICIDIN 硼替佐米 细胞凋亡 

分 类 号：R979.114[医药卫生—药品]