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


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

    Abstract:

    Objective:In our previous study,tumor necrosis factor (TNF)-α was showed to enhance the osteogenic differentiation of bone marrow derived mesenchymal stem cells (MSC). In this study,we aimed to investigate what effects of TNF-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-α for once (T+1 cohort ) or once a day for seven days (T+7 cohort),then the TNF-α pre-treated MSC was directly co-cultured with myeloma cell line H929 cells for four days. At the end of co-culture,H929 cells were harvested and tested for the number of colony-forming units (CFU),mRNA levels of POU5F1,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+1 and control cohorts. Results:TNF-α treatment did not cause any evidently morphological alterations on bone marrow MSCs. When compared with the control and T+1 cohort,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.

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徐欣欣,夏 雷,杨 娇,汤 郁,陆 化,朱 彦,费小明.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[J].南京医科大学学报(自然科学版),2015,(2):178

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  • 收稿日期:2014-05-19
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  • 在线发布日期: 2015-02-13
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