青蒿琥酯联合来那度胺对多发性骨髓瘤耐药细胞株的作用

目的 探讨青蒿琥酯联合来那度胺对多发性骨髓瘤耐药(MM.1R)细胞株增殖的影响并分析其作用机制.方法 实验设4组,空白对照组只加RPMI 1640培养液、来那度胺组加20μmol/L来那度胺、青蒿琥酯组加25μg/mL青蒿琥酯、来那度胺联合青蒿琥酯组加20μmol/L来那度胺浓度和25μg/mL青蒿琥酯浓度,终体积为200μL,分别对MM.1R细胞株处理24和48 h,MTT法检测细胞增殖;Western blot检测来那度胺联合青蒿琥酯处理24和48 h,MM.1R细胞株NF-κB P65蛋白,Bcl-2家族抗凋亡蛋白Bcl-xl、Mcl-1、p-Bad,多药耐药基因MDR1蛋白表达产物P-gp的表达水平.结果 来那度胺、青蒿琥酯、来那度胺联合青蒿琥酯处理24和48 h,各组抑制率分别为15.16%、28.28%、45.98%和17.19%、34.53%、65.16%,与对照组比较差异均有统计学意义(P<0.05,P<0.01),来那度胺联合青蒿琥酯组抑制率高于单药处理组(P<0.05),两药联合有良好的协同效应,两药联合处理48 h抑制率高于24 h,抑制效应呈时间依赖性;来那度胺联合青蒿琥酯处理24和48 h,MM.1R细胞株P-gp、NF-κB P65、Bcl-xl、Mcl-1、p-Bad表达量均低于对照组(P<0.05),两药联合处理48 h,MM.1R细胞株NF-κB P65、Bcl-xl、Mcl-1、p-Bad蛋白表达量低于24 h(P<0.05),表现出时间依赖性.结论 青蒿琥酯、来那度胺可抑制耐药MM细胞增殖,两药联合有协同效应,时间依赖性下调NF-κB P65蛋白、Bcl-2家族抗凋亡蛋白Bcl-xl、Mcl-1、p-Bad的表达,降低P-gp表达水平可能为其逆转肿瘤细胞耐药的作用机制.

The study on combination effect of artesunate and lenalidomide on the treatment of bortezomib-resistant multi-ple myeloma cell line

Objective To investigate the combination effect of artesunate and lenalidomide on the proliferation of bortezomib-resistant multiple myeloma cell line MM .1R, and its molecular mechanism .Methods MM.1R cells were cultured in RPMI 1640 and treated with DMSO (control group), lenalidomide (20 μmol/L), artesunate (25 μg/mL), or lenalidomide (20μmol/L) plus artesunate (25μg/mL), separately.After 24 or 48 h, the cell proliferation rates were assessed by MTT assay, and the NF-κB (P65), Bcl-xl, Mcl-1, p-Bad, and P-gp protein levels were measured by Western blot .Results The inhibitory rate of cell proliferation was 15.16%by lenalidomide for 24 h, 28.28%by lena-lidomide for 48 h, 45.98%by artesunate for 24 h, 17.19%by artesunate for 24 h, 34.53%by lenalidomide plus artesu-nate for 24 h, or 65.16%by lenalidomide plus artesunate for 48 h (P<0.05).NF-κB (P65), Bcl-xl, Mcl-1, p-Bad, and P-gp proteins were down -regulated by lenalidomide plus artesunate treatment in a time -dependent manner . Conclusion Both artesunate and lenalidomide inhibit MM .1R cell proliferation .Combination of artesunate with lenalido-mide can enlarge this effect through down -regulating NF-κB (P65), Bcl-xl, Mcl-1, p-Bad, and P-gp.