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目的 :多发性骨髓瘤(multiple myeloma,MM)表现有骨破坏和骨髓微环境的异常。近来发现MM患者的骨髓间充质干细胞(mesenchymal stem cell,MSC)有多种异常。既往研究发现经肿瘤坏死因子(tumor necrosis factor,TNF)-α预处理的骨髓MSC成骨分化潜能增强。本实验进一步研究TNF-α预处理的骨髓与骨髓瘤细胞株H929共培养后,对H929的影响。方法 :骨髓MSC经TNF-α单次(T+1组)或多次预处理(T+7组)后,与H929直接共培养3 d后,收集H929细胞分别检测其集落形成能力,定量RT-PCR检测POU5F1、SOX2和NANOG基因,以及微小RAN(miRNA)-15a/16的表达水平,并与对照组比较。结果:各组H929细胞均有POU5F1、SOX2、NANOG和miRNA-15a/16的检出。与对照组和T+1组相比较,T+7组的POU5F1和SOX2基因的表达水平下降且有统计学意义(P<0.05);miRNA-15a/16表达水平均上升且有统计学意义(P<0.05);CFU数减少且有统计学意义(P<0.05)。结论:经TNF-α多次预处理的骨髓MSC与MM细胞相互作用后,抑制MM细胞作用更加明显,具体机制有待进一步研究。
OBJECTIVE: Multiple myeloma (MM) exhibits bone destruction and abnormalities in the bone marrow microenvironment. Recently, it has been found that there are many abnormalities in mesenchymal stem cells (MSCs) of MM patients. Previous studies found that bone marrow MSCs pretreated with tumor necrosis factor (TNF) -α enhanced osteogenic differentiation potential. This experiment further study of TNF-α pretreatment of bone marrow and myeloma H929 co-culture, the impact of H929. Methods: After co-cultured with H929 for 3 days, the bone marrow MSCs were harvested (T + 1 group) or multiple pretreatments (T + 7 group) for 3 days. H929 cells were harvested for colony formation assay. Quantitative RT The expression levels of POU5F1, SOX2 and NANOG genes, and minute RAN (miRNA) -15a / 16 were detected by PCR and compared with the control group. Results: The detection of POU5F1, SOX2, NANOG and miRNA-15a / 16 in H929 cells of all groups. Compared with the control group and the T + 1 group, the expression levels of POU5F1 and SOX2 in T + 7 group decreased significantly (P <0.05), and the expression levels of miRNA-15a / 16 increased P <0.05). The number of CFU decreased and was statistically significant (P <0.05). CONCLUSION: After MMT pretreated by TNF-α for several times, the interaction between MM cells and MM cells is more obvious. The specific mechanism remains to be further studied.