摘要
目的研究姜黄素与马法兰联用对人多发性骨髓瘤耐药细胞株MOLP-2／R的耐药逆转作用及其与FA／BRCA途径可能存在的关系，进一步探讨其作用机制。方法采用MTT法检测细胞增殖抑制率；采用Western blot方法检测FA／BRCA途径中的关键蛋白FANCD2单泛素化表达；采用流式细胞术测定细胞周期、细胞内药物浓度及细胞凋亡率。结果姜黄素可增强马法兰对多发性骨髓瘤耐药细胞株MOLP-2／R的毒性，马法兰的阳。值由44．5μmol／L降至19．0μmol／L。该作用是通过抑制FA／BRCA途径中的关键蛋白FANCD2单泛素化来实现的。姜黄素和马法兰联合作用组与单用马法兰组相比，MOLP-2／R细胞的凋亡率由（23．3±0．6）％增至（52．6±0．8）％，G2／M期细胞由9．1％增至18．5％，细胞内药物荧光强度由15．2±0．3增至21．4±0．8。而姜黄素单药作用则无此效应，也不伴有FA／BRCA途径受抑制。结论姜黄素与小剂量马法兰合用可产生协同作用，姜黄素通过抑制FA／BRCA途径中的FANCD2单泛素化来逆转MOLP-2／R细胞对马法兰的耐药性。姜黄素介导的FA／BRCA途径受抑制，能增强马法兰对MOLP-2／R细胞的凋亡诱导及G2／M期阻滞作用，提高细胞内药物浓度。
Objective To investigate the reverse effect of mutidrug resistance of curcumin combined with melphalan on the mutidrug-resistant human multiple myeloma cell line MOLP-2/R and the relation with FA/BRCA pathway. Methods The inhibitory effects of the drugs on the growth of MOLP-2/R cells were determined by MTT assay. Cell cycle analysis, intracellular drug concentration and apoptosis were assayed by flow cytometry. The expression of FANCD2 monoubiquitination was determined by Western blot analysis. Resuits Co-administration of curcumin and melphalan had an synergistic inhibitory effects on the proliferation, IC50 of melphalan with 10μmol/L cureumin reduced from 45.5 μmol/L to 19 μmol/L in MOLP-2/R cells. The apoptosis percentage of MOLP-2/R cells was significantly increased from （23.3±0.6）% to （52.6% ±0.8 ） % by the treatment of melphalan 20 μmol/L plus cureumin 10 μmol/L with the increased percentage of cells in the G2/M phase （from 9.1% to 18.5 % ） and enhanced intraeellular drug concentration of MOLP-2/ R cells（ from 15.2 ±0.3 to 21.4 ±0.8 ）. The effects were accompanied with inhibition of FA/BRCA pathway by down regulation of FANCD2 protein monoubiquitination. Conclusion Curcumin combined with melphalan results in synergistic effects and reverses multiple drug resistance of MOLP-2/R cells effectively. The inhibition of FA/BRCA pathway may be the mechanism.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2009年第1期33-37,共5页
Chinese Journal of Hematology
