肿瘤坏死因子预处理的骨髓间充质干细胞对骨髓瘤细胞株H929集落形成能力及miRNA-15a/16等相关基因表达的影响  

Bone marrow derived mesenchymal stem cells co-cultured with tumor necrosis factor suppress myeloma cell line H929 malignant behavior

作　　者：徐欣欣[1] 夏雷[1] 杨娇[1] 汤郁 陆化[3] 朱彦[1] 费小明[1] 

机构地区：[1]江苏大学附属医院血液科 [2]风湿科,江苏镇江212001 [3]南京医科大学第一附属医院血液科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2015年第2期178-182,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81202358);镇江市社会发展项目(SH2011021)

摘　　要：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：In our previous study ,tumor necrosis factor （TNF）-a was showed to enhance the osteogenie difterentiatiun uf bone marrow derived mesenchymal stem cells （MSC）. In this study,we aimed to investigate what effects of TN F-α pre-treated bone marrow MSC on myeloma cells when they were in vitro co-cultured. Methods：Normal bone marrow MSC was treated with TNF-c~ [or once（T＋l cohort ） or once a day for seven days（T＋7 cohort）,then the TNF-α pre-treated MSC was directly co-cuhured with myeloma cell line H929 cells for tour days. At the end of co-culture,H929 cells were harvested and tested for the number of eolony-torming units （CFU） ,mRNA levels of POU5FI, SOX2 and NANOG and miroRNA levels of miRNA-15a/16 by real-time PCR. The results from T＋7 cohort were statistically compared with both T＋I and control cohorts. Results：TNF-a treatment did not cause any evidently morphological alterations on hone marrow MSCs. When compared with the control and T＋l eohort,our data showed the number of CFU in H929 was statistically decreased in T＋7 cohort,the mRAN levels of POU5F1 and SOX2 were statistically down-regulated in T＋7 cohort as well. However,the microRAN levels of both miRNA-15a and miRNA-16 were statistically up-regulated. Conclusion： Multiple TNF-α pre-treated bone marrow MSC was showed to has inhibitory effects on MM cells.

关 键 词：多发性骨髓瘤 间充质干细胞 TNF-α 骨髓微环境 

分 类 号：R733.3[医药卫生—肿瘤]