HMGB1基因沉默对阿霉素诱导K562/A02细胞凋亡增敏作用的研究  被引量:7

Enhancive effect of HMGB1 gene silence on adriamycin-induced apoptosis in K562/A02 drug resistance leukemia cells

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作  者:谢岷[1] 康睿[1] 俞燕[1] 朱珊[2] 贺钰磊[1] 许望琼[1] 唐道林[1] 曹励之[1] 

机构地区:[1]中南大学湘雅医院儿科,长沙410008 [2]中南大学湘雅医学院病理生理学系

出  处:《中华血液学杂志》2008年第8期549-552,共4页Chinese Journal of Hematology

基  金:国家自然科学基金(30571982、30772353)

摘  要:目的探讨高迁移率族蛋白1(HMGB1)基因沉默对白血病细胞耐药逆转的作用。方法将HMGB1基因特异性干扰RNA(HMGB1 siRNA)导入K562/A02细胞中,通过Western blot和RT—PCR方法检测HMGB1基因在转染前后的表达;WST8法检测阿霉素(ADM)对转染前后K562/A02细胞的半数抑制浓度(IC50);流式细胞术检测凋亡细胞百分率;Western blot检测线粒体促凋亡蛋白Smac/DIABLO的释放:采用caspase活性定量检测试剂盒分析caspase-3的活性。结果①与未经处理的K562/A02细胞组相比,转染HMGB1 siRNA的K562/A02细胞组HMGB1 mRNA和蛋白水平分别下降86%和71%;②HMGB1基因沉默使K562/A02细胞对ADM的药物敏感性增强,其IC50值从转染前的(4.83±0.08)μg/ml降低到(1.33±0.10)μg/ml,并在ADM浓度为1μg/ml和5μg/ml时,细胞凋亡百分率分别增加27%、32%;③HMGB1基因沉默可促进ADM所致Smac/DIABLO从线粒体向胞浆释放。并增加caspase-3的活性。结论HMGB1基因沉默能明显增加K562/A02细胞对ADM的敏感性,逆转K562/A02细胞对ADM耐药。Objective To investigate the effect of high mobility group boxl ( HMGB1 ) gene silence on adriamycin (ADM)-induced apoptosis in K562/A02 drug resistance leukemia cells. Methods K562/ A02 cells were transient transfected with HMGB1- small interference RNA(siRNA) vector, and the levels of HMGB1 gene differential expression pre-and post-transfection were measured by RT-PCR and Western blotting. 50% inhibition concentration (IC50) of ADM on K562/A02 was determined by WST-8 assay. Cell apoptosis was assessed by flow cytometry. The release of Smac/DIABLO from the mitochondria to the cytoplasm was assayed by Western blotting. Activity of Caspase-3 was assayed with a Caspase Colorimetric Assay Kit. Results ①The HMGB1 expression at mRNA and protein levels in HMGB1 siRNA transfected K562/A02 cells were decreased by 86% and 71% respectively compared with control. ②Suppression of HMGB1 by siRNA in K562/A02 cells resulted in a reversal of the resistance to ADM, and decreased IC50 from (4.83 ± 0.08 ) μg/ml to ( 1.33 ± 0.10 ) μg/ml. 1 μg/ml and 5 μg/ml of ADM treatment increased cell apoptotic rate by 27% and 32% respectively.③HMGB1 suppression in K562/A02 cells significantly promoted ADM- induced Smac/DIABLO release from the mitochondria to the cytoplasm, and increased the activities of Caspase-3. Conclusion HMGB1 gene silence can enhance sensitivity of K562/A02 cells to ADM and reverse cell resistant to ADM.

关 键 词:基因 HMGB1 细胞系 K562/A02 细胞凋亡 抗药性 多药 RNA干扰 

分 类 号:R686[医药卫生—骨科学]

 

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