姜黄素对多发性骨髓瘤细胞株MOLP-2/R的耐药逆转作用及与FA/BRCA途径的关系

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IC50值由44.5 μmoL/L降至19.0 μmol/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.

Reversal of mutidrug resistance of the drug resistant human multiple myeloma cell line MOLP-2/R by curcumin and its relation with FA/BRCA pathway

Objective To investigate the reverse effect of mutidrug resistance of cureumin 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 de-termined by MTT assay. Cell cycle analysis, intracellular drug concentration and apeptosis were assayed by flow cytometry. The expression of FANCD2 monoubiquitination was determined by Western blot analysis. Re-sults Co-administration of curcumin and melphalan had an synergistic inhibitory effects on the proliferation, IC50 of melphalan with 10 μmol/L curcumin 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 curcumin 10 μmol/L with the increased percentage of cells in the G2/M phase (from 9.1% to 18.5%) and enhanced intracellular 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 path-way by down regulation of FANCD2 protein monoubiquitination. Conclusion Curcumin combined with mel-phalan 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.