核转录因子RelB对慢性B淋巴细胞白血病细胞蛋白酶体抑制剂敏感性的影响  

Effect of nuclear transcription factor RelB on the proteasome inhibitor-sensitivity of chronic lymphocytic leukemia cells

作　　者：徐晶晶[1] 周鹏[1] 孙爱宁[2] 国风[1] 

机构地区：[1]苏州大学附属第一医院中心实验室,215006 [2]苏州大学附属第一医院血液科,215006

出　　处：《中华血液学杂志》2014年第6期524-527,共4页Chinese Journal of Hematology

基　　金：国家自然科学基金（81172433、81070405）;江苏省自然科学基金（BK2011306）;苏州市科技发展计划（SYS201337）

摘　　要：目的 探讨核转录因子RelB对慢性B淋巴细胞白血病（B-CLL）细胞蛋白酶体抑制剂敏感性的影响.方法 以初诊B-CLL患者CD5＋CD19＋细胞为研究对象,用RT-PCR法检测NF-κB家族成员RelB mRNA的表达水平;采用凝胶迁移实验及TransAMTMELISA法分析细胞核中RelB的活性;根据CLL细胞RelB活性将细胞分为RelB阳性（RelB＋）和RelB阴性（RelB-）两组,与人骨髓基质细胞共培养,分别加入氟达拉滨（10 μmol/L）、MG-132（1 μmol/L）和硼替佐米（1μmol/L）作用不同时间,采用碘化丙锭染色法分析药物对RelB＋和RelB-细胞死亡率的影响.结果 CLL细胞存在RelB mRNA表达及RelB活化,不同CLL细胞RelB mRNA表达水平及RelB活化水平不一.与骨髓基质细胞共培养时,三种药物均可促进细胞死亡,并具有时间依赖性.氟达拉滨对RelB＋和RelB-组CLL细胞的杀伤活性无明显差异,作用72 h后,两组CLL细胞死亡率分别为（61.11±6.91）％和（67.57±9.45）％;MG-132处理72 h对RelB＋细胞的杀伤活性高于RelB-组,两组细胞死亡率分别为（66.22±3.39）％和（51.07±5.93）％;硼替佐米处理24及48 h对RelB＋细胞杀伤活性高于RelB-组.24 h,两组细胞死亡率分别为（75.50±4.66）％和（66.32±10.20）％;48 h,死亡率分别为（92.11±3.14）％和（85.84±5.81）％.结论 骨髓来源CLL细胞中存在以RelB为代表的非经典性NF-κB活性.RelB活性增强可增加CLL细胞对蛋白酶体抑制剂硼替佐米和MG-132的敏感性,而对氟达拉滨的敏感性无明显影响.Objective To investigate the effect of nuclear transcription factor RelB on the proteasome inhibitor-sensitivity of chronic lymphocytic leukemia （CLL） cells.Methods The mRNA expression of RelB in CD5 ＋ CD19 ＋ CLL cells from BM was analyzed by reverse transcription PCR （RT-PCR）.The RelB activity was examined by electromobility shift assay （EMSA） and an ELISA-based NF-κd3 family transcription factor activity assay.CLL cells were classified into RelB＋ and RelB-groups according to RelB activity.The frequencies of cell death of CLL cells cultured with human bone marrow stromal cells （hBMSCs） after treatment with PS-341,MG-132 or fludarabine were determined by PI staining.Results RelB mRNA expression and RelB activity could be detected in CLL cells at variable levels.Fludarabine （10 μmol/L）,MG-132 （1 μmol/L） and PS-341 （1 μmol/L） could induce cell death of RelB＋ and RelB-CLL cells co-cultured with hBMSCs in a time dependent manner.There was no significant difference in the fludarabine sensitivity between RelB ＋ and RelB-CLL cells,and the frequencies of cell death of RelB＋ and RelB-CLL cells were （61.11 ±6.91）% and （67.57±9.45）%,respectively,when treated with fludarabine for 72 h.RelB＋ CLL cells were more sensitive to MG-132 than RelB-CLL cells for 72 h,and the frequencies of cell death were （66.22±3.39） % and （51.07±5.93） %,respectively.RelB＋ CLL cells were more sensitive to PS-341 than RelB CLL cells for 24 and 48 h treatment,and the frequencies of cell death of RelB＋ and RelB-CLL cells were （75.50±4.66）% and （66.32±10.20）% for 24 h,（92.11±3.14）% and （85.84 ± 5.81）% for 48 h treatment,respectively.Conclusion The alternative NF-κB activity was detected in bone marrow derived CLL cells.Enhancement of RelB activity may increase CLL cells＇ sensitivity to proteasome inhibitor bortezomib and MG-132.However,the sensitivity of CLL cells to fludarabine had no relationship to RelB activity.

关 键 词：白血病 淋巴细胞 慢性 B细胞 转录因子RelB 蛋白酶体抑制剂 

分 类 号：R733.72[医药卫生—肿瘤]