复方当归注射液逆转K562/A02细胞多药耐药的研究

目的:研究复方当归注射液对人红白血病多药耐药(MDR)细胞株K562/A02MDR1、多药耐药相关蛋白(MRP)、拓扑异构酶Ⅱ(TopoⅡ)、谷光甘肽-S-转移酶-π(GST-π)基因及其相应蛋白表达的影响。方法:采用台盼蓝染色法检测复方当归注射液的细胞毒作用;采用半定量逆转录聚合酶链反应(RT-PCR)和免疫印迹法分别检测复方当归注射液在非细胞毒浓度(IC10)作用下对K562/A02细胞上述基因及其表达的影响。结果:复方当归注射液作用后,K562/A02细胞中MDR1基因及P-gp表达降低(P<0.01);TopoⅡ蛋白表达增高(P<0.01),TopoⅡ基因表达无显著变化(P>0.05);MRP、GST-π基因及其蛋白的表达无明显变化(P>0.05)。结论:复方当归注射液能部分逆转K562/A02细胞的耐药性,其作用机制可能与下调K562/A02细胞MDR1mRNA的表达,导致细胞膜上P-gp的表达量减少及在蛋白水平上调K562/A02细胞TopoⅡ的表达有关。

Reversal of Multidrug Resistance in K562/A02 Cells by Compound Angelica sinensis Injection

OBJECTIVE： To study the effects of Compound Angelica sinensis injection on the protein expression and gene of human erythroleukemia multidrug resistance （MDR） K562/A02 cell line, such as MDR1 gene, multidrug resistance-associated protein （MRP）, topoisomerase Ⅱ （Topo Ⅱ） and glutathione-S-Transferase-π（GST-π）. METHODS： Trypan blue staining was applied to measure the toxic effect of Compound A.sinensis injection. Semi-Quantitative RT-PCR and Western blot assay were applied to analyze the effect of Compound A.sinensis injection with condition of IC10 of ADM on above genes and their expression. RESULTS： After treating with Compound A.sinensis injection, mRNA expression of MDRI gene and P-gp were reduced while protein expression of Topo Ⅱ increased significantly （P〈0.01） ; the mRNA expression of Topo 11 and other gene had no conspicuous change （P〉0.05）. CONCLUSION： The multidrug resistance of K562/A02 cells partly reversed by Compound A.sinensis injection, which may associate with reduction of P-gp expression resulted by down-regulating mRNA expression of MDR1 gene and up-regulation of Topo Ⅱ expression.