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目的观察莪术醇对体外多发性骨髓瘤(multiple myeloma,MM)细胞生物学行为的影响,为莪术醇临床治疗MM提供理论依据。方法以骨髓间充质干细胞(bone marrow derived mesenchymal stem cells,BMSCs)和骨髓瘤8226细胞株(multiple myeloma cell line 8226,RPMI 8226)为研究对象,分为RMPI 8226单独培养组(RMPI 8226)和BMSCs RMPI 8226细胞共培养组(BMSCs+RMPI 8226),分别加入不同浓度(0.1、0.5、1、10μg/m L)的莪术醇,采用流式细胞术检测莪术醇对各组RPMI 8226增殖、周期及凋亡的影响;应用逆转录聚合酶链反应(reverse transcriptase-polymerase chain reaction,RT-PCR)检测BMSCs成骨分化基因核因子κb受体活化因子配体(receptor activator of nuclear factorκb ligand,RANKL)及骨保护素(osteoprotegerin,OPG)的表达水平。结果莪术醇可诱导RMPI 8226细胞周期阻滞,且对单独培养组RMPI 8226细胞的周期阻滞明显高于BMSCs共培养组;莪术醇能明显抑制RPMI 8226细胞增殖,诱导RPMI 8226细胞凋亡(均P<0.01),BMSCs可明显降低莪术醇诱导的RPMI 8226细胞凋亡;莪术醇可下调骨髓瘤骨病相关基因RANKL表达,上调OPG表达。结论莪术醇能干扰MM细胞周期,诱导细胞凋亡,在下调RANKL表达的同时,上调OPG表达;BMSCs共培养明显抑制莪术醇诱导的MM细胞凋亡,可能与MM细胞耐药有关。
Objective To observe the effect of curcumol on the biological behavior of multiple myeloma (MM) in vitro and to provide a theoretical basis for the clinical treatment of MM with curcumol. Methods Bone marrow derived mesenchymal stem cells (BMSCs) and myeloma cell line 8226 (RPMI 8226) were divided into RMPI 8226 group (RMPI 8226) and BMSCs (BMSCs + RMPI 8226) group were treated with curcumol at different concentrations (0.1, 0.5, 1 and 10 μg / mL) respectively. Flow cytometry was used to detect the proliferation, (RT-PCR) were used to detect the expression of receptor activator of nuclear factor kappa B ligand (RANKL) and Osteoprotegerin (osteoprotegerin, OPG) expression levels. Results Curcumol induced cell cycle arrest of RMPI 8226 cells, and the cell cycle arrest in RMPI 8226 cells was significantly higher than that of BMSCs co-cultured with curcumol. Curcumol significantly inhibited the proliferation of RPMI 8226 cells and induced the apoptosis of RPMI 8226 cells P <0.01). BMSCs significantly decreased the apoptosis of RPMI 8226 cells induced by curcumol. Curcumol could down-regulate the expression of RANKL and up-regulate the expression of OPG in osteosarcoma of bone marrow. Conclusion Curcumol can interfere with the cell cycle of MM and induce apoptosis. While down-regulating the expression of RANKL, up-regulating the expression of OPG. Co-culture of BMSCs significantly inhibits the apoptosis of MM cells induced by Curcumol, which may be related to the resistance of MM cells.