中国药理学与毒理学杂志2013，Vol.27Issue(1)：77-83,7.DOI:10.3867/j.issn.1000-3002.2013.01.015
米哚妥林对P糖蛋白介导的多药耐药的逆转作用
Reversal effect of midostaurin on P-glycoprotein-mediated multidrug resistance
摘要
Abstract
OBJECTIVE To determine the potency of midostaurin to sensitize P-glycoprotein (P-gp)-mediated multidrug resistance. METHODS K562/A02 or K562 cells were treated with midostaurin 0.5, 1 and 5μmol·L-1 for 72 h, and determined by MTS assay to evaluate the cytotoxicity effect of midostaurin in sensitive and resistant cells. The effect of inhibitors on drug resistance was accessed by exposing cells to a low concentration ( ≤ IC10) of anticancer drugs such as doxorubicin, vincristine and paclitaxel in the absence or presence of midostaurin 0.25 and 0.5 μmol·L-1 followed by MTS assay. Rh-123 accumulation was detected by FACS in K562/A02 or K562 cells following 30 min incubation in the absence or presence of midostaurin or verapamil 0.5 and 10 μmol·L-1. The effect of midostaurin on the protein and mRNA expression level was determined separately by Western blotting and qPCR. Midostaurin was firstly incubated with P-gp rich crude membranes and MgATP at 37℃ for 40 min before luminescence was initiated by ATP detection buffer, and then the remaining ATP was detected after incubation at room temperature for 40 min. RESULTS Midostaurin showed equal cytotoxicity in both K562/A02 and K562 cells , and the cell survival rate reached 80% after midostaurin 0.5 μmol·L-1 for 72 h. Midostaurin 0.5 μmol·L-1 significantly potentiated the cytotoxicity of doxorubicin, paclitaxel and vincristine to K562/A02 rather than to P-gp negative parental cell K562. Midostaurin 0. 5 and 10 μmol·L-1 remarkably increased the accumulation of Rh-123 in K562/A02 cells and had better effect than verapamil under the same condition, but had no effect on sensitive cells. However, the ATPase activity assay showed that P-gp-mediated ATP hydrolysis could be inhibited by midostaurin. Midostaurin 0.5 μmol·L-1 for 72 h did not influence the expression of P-gp at the protein and mRNA level, and had little impact on the total and phosphorylated forms of AKT and ERK1/2. CONCLUSION Midostaurin reverses multidrug resistance by inhibiting the efflux function of P-gp, unlikely to be a substrate of P-gp.关键词
米哚妥林/多药耐药相关蛋白/糖蛋白类/蛋白激酶抑制剂Key words
midostaurin/ multidrug resistance-associated proteins/ glycoproteins/ protein kinase inhibitors分类
医药卫生引用本文复制引用
魏寅祥,赵青,任志广,贾砚寒,李新颖,黎燕,彭晖,马远方..米哚妥林对P糖蛋白介导的多药耐药的逆转作用[J].中国药理学与毒理学杂志,2013,27(1):77-83,7.基金项目
国家自然科学基金(30873083) （30873083）
国家自然科学基金(811730782) （811730782）
