MG132诱导HL-60细胞表达共刺激分子CD86及对混合淋巴细胞反应的影响  

Role of MG132,a proteasome inhibitor,in inducing expression of costimulatory molecules CD86 in leukemic cells and its effect on allogeneic mixed lymphocyte reaction

作　　者：周咏明[1] 郭伟[2] 薛克营[3] 周浩[1] 程艳香[4] 郭天南[1] 李慧玉[1] 黄士昂[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院干细胞应用与研究中心,武汉430022 [2]华中科技大学同济医学院附属同济医院内分泌科 [3]华中科技大学同济医学院附属同济医院呼吸科 [4]华中科技大学同济医学院附属同济医院妇产科

出　　处：《中华医学杂志》2007年第10期710-713,共4页National Medical Journal of China

基　　金：国家杰出青年基金(30225038);国家科技部973项目资助项目(00CB510103)

摘　　要：目的研究蛋白酶体抑制剂 MG132诱导 HL-60细胞表达共刺激分子 CD80、CD86及其对混合淋巴细胞反应的作用。方法流式细胞仪检测 MG132诱导 HL-60细胞表达共刺激分子CD80、CD86及细胞活力;逆转录聚合酶链反应(RT-PCR)分析 CD80和 CD86 mRNA 表达情况;75 Gy照射 MG132诱导的 HL-60细胞后,用 CCK-8检测提高共刺激分子表达后,HL-60细胞对健康人单个核细胞的增殖作用。结果 MG132上调 HL-60细胞表达 CD86,高浓度的 MG132对 HL-60细胞有直接杀灭作用,MG132诱导 HL-60细胞表达共刺激分子 CD86后,对健康人单个核细胞起增殖作用。结论 MG132诱导 HL-60细胞表达共刺激分子 CD86,促进对健康人单个核细胞的增殖作用。Objective To investigate the role of proteasome inhibitors MG132 in the inducing the expression of the costimulatory molecules CD80 and CD86 in leukemia ceils and its effect on allogeneic mixed lymphocyte reaction. Methods Acute myelocytic leukemia ceils of the line HL-60 and chronic myelocytic leukemia ceils of the line K562 were cultured. 7-AAD staining and flow cytometry （ FC ） were used to examine the viability of the cells. MG132, a proteasome inhibitor, of the concentrations of 2 or 3 μmol/L was added into the culture fluid of HL-60 cells for 24 h and 48 h respectively and then annexin V/ 7-AAD staining and FC were used to detect the apoptosis of the cells. HL-60 and K562 cells treated with 1 p.mol/L MG132 for 24 h and 48 h respectively, anti-CD80 and anti-CD86 antibodies were added, then FC was used to detect the expression of CD80 and CD86. The mRNA expression of CD86 in the HL-60 cells treated with 1 μmol/L MG132 was examined by RT-PCR. HL-60 and K562 cells were treated by 1 μmol/L MG132 for 48 h and then underwent irradiation of 75 Gy Co-60 to kill the cells with their antigenicity preserved. Peripheral blood mononuclear cells （PBMNC） of healthy volunteers, as reactive cells, were isolated and inoculated into the Co-60 treated HL-60 and K532 cells of different concentrations, as stimulating cells, for 5 d, CCK-8, a new agent to detect the cell viability, was added for 4 h, and then the A value of absorbance was measured at the wave length of 450 nm of enzyme labeling instrument. Control groups were set up for all tests. Results The cell viability rates of the HL-60 cell treated with 1 p.mol/L MG132 for 24 h and 48 h were 92.95% and 85.87% respectively. The apoptotic rats of the HL-60 cells treated with MG132 were increased dose- and time-dependently. Before MG132 treatment K562 cells did not express CD86, and the CD86 expression of the HL-60 cells was up-regulated time-dependently （ all P 〈 0.01 ）. The mRNA expression of CD86 in the HL-60 treated with MG132 was up-regulated time-depende

关 键 词：蛋白酶抑制药 抗原 CD86 淋巴细胞培养试验 混合 

分 类 号：R73-3[医药卫生—肿瘤]