骨髓瘤细胞可诱导骨髓间充质干细胞的基因表达谱发生暂时和 (或)长期性改变

目的:正常骨髓间充质干细胞(mesenchymal stem cell,MSC)在与骨髓瘤细胞相互作用过程中,MSC的全基因表达谱的改变目前尚未有报道,本研究就对此进行研究并进一步探索多发性骨髓瘤的发病机制。方法:正常人骨髓MSC与骨髓瘤细胞株在Transwell共培养体系培养前后,用全基因表达芯片检测MSC全基因mRNA的表达谱,比较正常MSC在与骨髓瘤细胞共培养(MC组)或共培养后去除骨髓瘤细胞后继续单独培养的MSC(MA组),和MSC单独培养的对照组(MK组)基因表达谱的变化。结果:MC组与MK组相比较,在所有分析的10 000个基因中共发现837个差异基因(837/10 000,8.37%),其中有472个基因表达上调(472/837,56.39%),365个基因表达下调(365/837,43.61%)。而MA组与MK组相比较,共发现367个差异基因,其中有218个基因表达上调(218/367,59.40%),149个基因表达下调(149/367,40.60%)。从芯片结果中筛选出MMP1、FGFR2、ANGPTL4、MFAP5、TGM2、STC1、CCL7和IL-32这8个基因,经定量PCR验证后的结果与基因芯片结果相一致。结论:骨髓瘤细胞可以诱导正常骨髓MSC多种基因表达改变,并且有些改变即使在骨髓MSC脱离骨髓瘤细胞后仍可存在;此外,初步筛选到8个差异表达基因,其中7个基因在多发性骨髓瘤发病机制中的作用既往未见报道,有待今后进一步研究。

Both transient and sustained gene expression profile alterations of bone marrow mesenchymal stem cells induced upon co-cultured with myeloma cells

Objective:Recent researches suggested that the abnormalities of bone marrow microenvironment have important implications for the genesis and evolution of multiple myeloma. However,the gene expression profile (GEP)changes in the interaction between bone marrow mesenchymal stem cell(MSC) and myeloma cells are not reported. Methods:To investigate the pathogenesis of multiple myeloma,cDNA microarray was employed to investigate the GEP of MSC before and after interacting with myeloma cells. And the genes with differential expression MSC co-cultured with myeloma cells (the MC groups)or cultured alone to continue after the removal of MM cells (MA groups)compared with the control groups (the MK groups)were screened by scanning and analyzed by computer software. Results:After co-cultured of MSC with MM cells (the MC groups),among total 10 000 genes,we filtered a total of 837 differential representing genes (837/10 000,8.37%),of which there were 472 up-regulated genes (472/837,56.39%)and 365 genes were down-regulated(365/837,43.61%). We found 367 differentially expressed genes in the MA groups compared to the control ones,including 218 up-regulated genes (218/367,59.40%)and 149 down-regulated genes(149/367,40.60%). Then we selected eight genes(MMP-1,FGFR2,ANGPTL4,MFAP5,TGM2,STC1,CCL7 and IL-32)from differential genes to verify by real-time quantitative PCR and their functions were explored. The Real-time quantitative PCR results were consistent with cDNA microarray results. Conclusion: It is indicated that myeloma cells induce a variety of gene expression changes of normal bone marrow MSC,and some changes still exist even after the removal of MM cells. In addition,among the eight selected genes in this study,seven of them have not been reported to be involved in the pathogenesis of multiple myeloma.