应用蛋白质芯片对汉防己甲素单用及与屈洛昔芬伍用逆转白血病细胞耐药机理的研究  

Using Protein Chips to Study Mechanism Underlying Reversion of Drug Resistance in Leukemia Cells in Tetrandrime alone or in Combination with Droloxifen

作　　者：陈宝安[1] 杜鹃[1] 张春秀[2] 程坚[1] 高峰[1] 陆祖宏[2] 

机构地区：[1]东南大学附属中大医院血液科 [2]东南大学生物科学与医学工程系,南京210009

出　　处：《中国实验血液学杂志》2005年第6期999-1003,共5页Journal of Experimental Hematology

摘　　要：本研究的目的是应用蛋白质芯片检测汉防己甲素(Tet)单独及与屈洛昔芬(Drol)伍用对作用的白血病细胞表面的耐药蛋白,包括P糖蛋白(Pgp)、多药耐药相关蛋白(MRP1)、乳腺癌耐药蛋白(BCRP)表达的作用,为逆转剂的临床应用提供理论依据。选择位于膜表面Pgp、MRP1、BCRP耐药蛋白及其相应的抗体为研究体系,制备蛋白芯片,直接对逆转剂作用12、24和48小时的K562/A02细胞进行检测。结果表明:Tet和Drol联合作用24小时时检测到Pgp表达下调(Tet+Drol组:85.27±3.095,对照组:93.67±2.748,P<0.05)。经逆转剂单独及联合作用K562/A02细胞48小时时均检测到Pgp表达下调,且联合应用两药对Pgp表达下调作用明显(Tet+Drol:82.62±3.227,Tet:86.44±2.906,Drol:87.23±2.049,对照组:93.67±2.748,P<0.05)。检测结果与流式细胞仪检测结果一致。结论:逆转剂Tet和Drol对K562/A02细胞的Pgp下调呈时间依赖性。联合作用24小时时出现下调Pgp表达,单独作用48小时均下调Pgp表达,联合用药时下调作用明显。不同检测时间均未见下调MRP1和BCRP的表达。The aim of this study was to investigate the mechanism to reverse the drug resistance of leukemia cells in tetrandrine （Tet） alone or in combination with droloxifen （Drol） by using protein chips and to lay the theoretical basis for the clinical applications. Three monoclonal antibodies against P-glycoprotein （P-gp）, the mutidrug resistanceassociated protein （ MRP1 ） and the breast cancer resistance protein （BCRP） were immobilized onto the agarose gel filmcoated glass slides. Protein chips were prepared respectively from K562/A02 cells cultured for 12, 24 and 48 hours with Tet alone or in combination with Drol. The results showed that Tet alone or in combination with Drol could decrease only the expression of P-gp in a time-dependent manner, the effect for 48 hours as follows ： Tet ＋ Drol 82. 620 ± 3. 227 ; Tet alone 86. 40 ± 2. 906 ; Drol alone 87. 230 ± 2. 049 ; control 93. 670 ± 2. 748 （ P 〈 0.05 ）. However, down-regulation of P-gp by K562/A02 cells cultured with Tet alone or in combination with Drol began at 24 hours （ Tet ＋ Drol 85. 270 ± 3. 095 ; control 93. 670 ± 2. 48, P 〈 0.05 ）. The results were coincident with that of FCM. It is concluded that Tet and Drol can downregulate the expression of P-gp in the time-dependent way. There is a significant difference between Tet alone and Tet combined with Drol at 24 hours （ P 〈 0.05 ）. The expression of MRP1 and BCRP are not closely correlated with the reversal mechanism of Tet and Drol, and which may be involved in the mechanism of this combination to reverse multidrug resistance in leukemia.

关 键 词：多药耐药 汉防己甲素 屈洛昔芬 蛋白质芯片 K562/A02 

分 类 号：R733.7[医药卫生—肿瘤] R979.1[医药卫生—临床医学]