谷胱甘肽含量对肿瘤多药耐药相关蛋白1的影响  

Effect of the content of glutathione for multidrug resistance-associated protein 1

作　　者：王涛[1] 马梁明[1] 牛燕燕[1] 任瑞瑞[1] 

机构地区：[1]山西医科大学附属山西大医院血液科,030032

出　　处：《肿瘤研究与临床》2014年第6期366-372,共7页Cancer Research and Clinic

基　　金：山西省青年科技研究基金（2010021035-3）

摘　　要：目的 研究应用丁硫氨酸亚砜胺（BSO）降低谷胱甘肽（GSH）含量后,三氧化二砷（As2O3）对K562/ADM细胞的诱导凋亡效应,及其对多药耐药相关蛋白1（MRP1）的抑制作用.方法 As2O3组（0.5、2.0、5.0 μmol/L）单独及联合100 μmol/L BSO作用于K562/ADM细胞,应用四甲基偶氮唑蓝（MTT）比色法检测K562/ADM细胞的增殖活性;Annexin V/PI标记法观察K562/ADM细胞的凋亡效应;分光光度法检测K562/ADM细胞内GSH含量变化;流式细胞术（FCM）检测的MRP1蛋白水平表达变化;反转录-聚合酶链反应（RT-PCR）方法检测MRP1的编码基因mRNA的表达变化.结果 降低GSH含量后,临床剂量As2O3（0.5、2.0 μmol/L）联合BSO 24 h内即可抑制K562/ADM细胞的增殖活性,诱导K562/ADM细胞发生凋亡,72 h单用As2O3（0.5、2.0 μmol/L）凋亡率为（8.32±2.11）％、（16.75±3.56）％,GSH含量降低后,两剂量组细胞的凋亡率分别为（82.15±9.28）％和（92.72±9.41）％.72 h后,临床剂量As2O3联合BSO抑制MRP1的荧光强度分别为8.20±0.92和10.40±2.33,明显低于单用高剂量As2O3组的荧光强度（21.30±3.09）;两药联合对于MRP1 mRNA的作用效果也明显强于单用高剂量As2O3组.结论 GSH的含量变化与As2O3的作用效果密切相关,As2O3联合BSO可有效诱导K562/ADM细胞发生凋亡,可有效抑制MRP1及其编码基因mRNA的表达.Objective To investigate the apoptosis-inducing effect,inhibiting multidrug resistanceassociated protein 1 （MRP-1） and mRNA expression of arsenic trioxide （As2O3） and buthionine sulfoximine （BSO） in multidrug-resistant cell K562/ADM.To compare the effect of As2O3 and the combined group.To determine the effect of intracellular glutathione （GSH） content on the arsenic effect.Methods The arsenic group （0.5 μmol/L,2.0 μmol/L,5.0 μmol/L） solo or combined BSO （100 μmol/L） applied in multidrugresistant cell K562/ADM.The cell proliferating activity was assessed with MTT assay.The cell apoptosis effect was detected by Annexin-V and propidium iodide （PI） staining.Intracellular GSH contents were measured using GSH Assay Kit by spectrophotometry.MRP1 expression was determined by flow cytometry.MRP1 mRNA expression were directed by semi-quantitative RT-PCR.Results With the GSH contents were degraded by the combination of clinic dose arsenic group （0.5 μmol/L,2 μmol/L） and BSO （100 μmol/L）,the K562/ADM cell proliferating activity was obviously inhibited and the cell apoptosis-inducing effect was increased in 24 hours.In 72 hours,the rate of apoptosis with arsenic （0.5 μmol/L,2 μmol/L） were （8.32± 2.11） %,（16.75±3.56） %.After the GSH contents were degraded,the rates of apoptosis in the combination group （clinic dose arsenic group） were （82.15±9.28） % and （92.72±9.41） %.The fluorescence intensity of MRP1 in 72 hours of the combination group （clinic dose arsenic group） was 8.20±0.92 and 10.40±2.33.The MRP1 attenuated effect of the combination group （clinic dose arsenic group） was obviously stronger than that of the high dose arsenic group （21.30±3.09）.Conclusions The intracellular GSH contents closely correlate with the arsenic effect.The cell apoptosis-inducing effect of the combination of clinic dose arsenic and BSO on K562/ADM cell is obvious increased.The combination of clinic dose arsenic and BSO obviously inhibit MRP1 expression and MRP1

关 键 词：肿瘤 多药耐药 三氧化二砷 谷胱甘肽 

分 类 号：R730.5[医药卫生—肿瘤]