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目的探讨蛋白质芯片在检测白血病细胞多药耐药(MDR)蛋白表达中的价值。方法以人红白血病细胞系K562及其耐药细胞系K562/A02为实验研究对象。将3种耐药蛋白P糖蛋白(P-gP)、多药耐药相关蛋白(MRP1)和乳腺癌耐药蛋白(BCRP)相应的单克隆抗体固定在玻片上形成微阵列,细胞直接与固定在芯片上的耐药抗体阵列反应,电荷耦合器件(CCD)检测反应结果,并与流式细胞术测定的结果进行比较分析。结果在K562细胞中,蛋白质芯片检测到P-gP和BCRP表达率低,MRP1有较高水平的表达;在K562/A02细胞中,P-gP和MRP1均有高水平表达,BCRP表达率低。流式细胞术结果显示,K562细胞P-gP、MRP1和BCRP表达率分别为5.98%±2.19%、95.80%±3.98%和1.03%±0.45%;K562/A02细胞P-gP、MRP1和BCRP表达率分别为92.67%±1.80%、97.18%±1.02%和3.98%±0.37%。经统计学分析,两种方法结果一致(Ρ>0.05)。结论利用蛋白质芯片检测MDR蛋白结果可靠,具有高通量、低成本、制备简单、测定快速的优点。
Objective To investigate the value of protein chip in the detection of multidrug resistance (MDR) protein in leukemia cells. Methods Human erythroleukemia cell line K562 and its drug-resistant cell line K562 / A02 were used as experimental subjects. Three monoclonal antibodies against P-gp, MRP1 and BCRP were immobilized on glass slides to form microarrays. The cells were directly and fixed Antibody-resistant antibody array reaction on chip, CCD (Charge Coupled Device) detection reaction results, and compared with the results of flow cytometry analysis. Results In K562 cells, the expression of P-gp and BCRP was low and the expression of MRP1 was higher in K562 / A02 cells. Both P-gp and MRP1 were highly expressed and the expression of BCRP was low in K562 / A02 cells. The results of flow cytometry showed that the expressions of P-gp, MRP1 and BCRP in K562 cells were 5.98% ± 2.19%, 95.80% ± 3.98% and 1.03% ± 0.45%, respectively. The expressions of P-gp, MRP1 and BCRP in K562 / A02 cells Rates were 92.67% ± 1.80%, 97.18% ± 1.02% and 3.98% ± 0.37%, respectively. After statistical analysis, the two methods are consistent (Ρ> 0.05). Conclusion The result of MDR protein detection by protein chip is reliable, with the advantages of high throughput, low cost, simple preparation and rapid determination.