Detection And Research Of Multidrug Resistance Proteins Based On Protein Array

Emergence of multidrug resistance is a major obstacle to successful chemotherapy of leukemia. One of the most common and well-studied mechanisms implicated in causing multidrug resistance are P-glycoprotein, the multidrug resistance-associated protein, and the breast cancer resistance protein, ATP-dependent, transmembrane drug efflux pump. Objective: To evaluate protein array in detection of mutidrug-resistance proteins and investigate the reversal mechanism of the Tetrandrine(Tet) and Droloxifen(Drol) alone or allied, and to provide theoretic evidence for the clinical application of them.Methods: The glass slides were modified with four different methods. K562, K562/A02 and clinic sample were used in the study. Three monoclonal antibodies-the P-glycoprotein (P-gP), the mutidrug resistance-associated protein (MRP1) and breast cancer resistance protein (BCRP) were immobilized onto the agarose film coated glass. Protein array was observed under a microscope and the image was captured with a CCD camera. The expression levels of three proteins were also measured with flow cytometry. Results: The glass slides were modified with agarose has the biggest surface area and highest capacity. The expression of P-gP and BCRP in K562 was very low, however, MRP1 was high. P-gP and MRP1 expressed highly in K562/A02, the expression of BCRP was low. Tet and Drol alone or allied can only decrease the expression of P-gP, which resulted in a time-dependent. The level of these proteins expression in relapsed-refractory group was higher than those in untreated and normal control groups. The method is coincident with FCM.Conclusion: The object of the research is to develop a new application of protein array, and to provide a novel method for cell detection which has the abilities of high throughput, high specificity, low sample consume, and low cost. The reversing mechanism of Tet and Drol has relation to decrease P-gP probably.