硼替佐米对多发性骨髓瘤细胞凋亡及分子伴侣BiP表达的影响  

Bortezomib-induced BiP Expression and Apoptosis in Multiple Myeloma Cells

作　　者：董红娟[1] 陈协群[1] 高广勋[1] 顾宏涛[1] 潘耀柱[1] 高瑛[1] 朱华锋[1] 

机构地区：[1]第四军医大学西京医院血液科陕西,西安710032

出　　处：《中国实验血液学杂志》2009年第1期107-110,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目;编号30871089

摘　　要：本研究旨在探讨蛋白酶体抑制剂硼替佐米对多发性骨髓瘤细胞株NCI-H929(H929)的凋亡诱导作用及其对H929细胞中分子伴侣BiP表达的影响。以不同浓度硼替佐米处理H929细胞24小时后,采用Annexin V-FITC/PI双染联合流式细胞术检测靶细胞凋亡率;用RT-PCR和Western blot检测靶细胞BiP mRNA和BiP蛋白的表达。结果表明:不同浓度硼替佐米(20、40、80nmol/L)均可诱导靶细胞凋亡,凋亡率分别为(15.73±0.67)%、(27.83±1.26)%及(44.17±2.25)%,呈剂量依赖性,且均显著高于未处理组(1.21±0.07%)(p<0.05);经不同浓度硼替佐米处理后,靶细胞BiP mRNA和BiP蛋白水平均高于未处理组,且二者变化一致;在相近的促凋亡条件下(凋亡率约40%-50%),标准内质网应激诱导剂brefeldin A(500ng/ml,24小时)对H929细胞BiP mRNA和BiP蛋白表达的上调作用与硼替佐米(80nmol/L,24小时)基本一致。结论:硼替佐米诱导H929细胞凋亡并伴有分子伴侣BiP mRNA和BiP蛋白表达上调,表明硼替佐米诱导的H929细胞凋亡极可能涉及内质网应激反应。This study was aimed to explore the effect of bortezomib on the apoptosis and expression of the molecular chaperon BiP in human multiple myeloma cell line NCI-H929 （H929）. After treatment of H929 cells with different concentrations of bortezomib for 24 hours, cell apoptosis was assayed by flow cytometry with Annexin V-HTC/PI staining, and the expression levels of BiP mRNA and protein were detected by RT-PCR and Western blotting analysis. The results showed that bortezomib of different concentrations （20, 40 and 80 nmol/L） induced apoptosis of H929 cells in dose-dependent manner, with apoptotic rates （ 15.73 ＋ 0. 67 ） %, （ 27. 83 ＋ 1.26 ） % and （ 44. 17 ＋ 2.25 ） % respectively, which were significantly higher than that in control （ 1.21 ＋ 0.07% ） （p 〈 0.05）. Bortezomib-induced upregulation of BiP mRNA levels was almost on a parallel with BiP protein when compared with control. Under the similar apoptosis-stimulating conditions with apoptotic rates varying from 40% to 50%, expression levels of BiP mRNA and BiP protein induced by the classical endoplasmic reticulum stressor Brefeldin A （500 ng/ml, 24 h） were almost consistent with those by bortezomib（80 nmol/L, 24 h）. It is concluded that bortezomib-induced apoptosis in/-/929 cells correlates closely with endoplasmic reticulum stress.

关 键 词：硼替佐米 骨髓瘤 NCI-H929细胞 BIP 内质网应激 细胞凋亡 

分 类 号：R733.3[医药卫生—肿瘤] R979.1[医药卫生—临床医学]