肿瘤坏死因子α对骨髓瘤患者骨髓间充质干细胞TAZ表达和成骨潜能的影响  

The effect of TNF-α on TAZ expression and osteogenic potential of mesenchymal stem cells from myeloma patients

作　　者：李炳宗 史明霞 李静[1] 马杰 陈磊 陈斌 赵春华 

机构地区：[1]中国医学科学院基础研究所组织工程研究中心,北京100730

出　　处：《中华肿瘤杂志》2009年第1期5-9,共5页Chinese Journal of Oncology

基　　金：国家高技术研究发展计划（2006AA02A109）;国家自然科学基金（30570771）.

摘　　要：目的探讨肿瘤坏死因子α（TNF-α）、具有PDZ结合域的转录共刺激因子TAZ和骨髓瘤骨病的关系。方法分离培养多发性骨髓瘤（MM）患者和正常人的骨髓间充质干细胞（MSCs），鉴定其生物学特性；酶联免疫吸附试验（ELISA）检测MM患者和正常人骨髓血清中TNF-α的浓度。MM患者骨髓CD138^＋细胞与正常人MSCs行Transwell成骨共培养后，采用实时定量PCR检测MSCs成骨指标和TAZ mRNA表达量，von Kossa染色法鉴定钙质沉积度。采用实时定量PCR和Western blot检测MSCs中TAZ mRNA和蛋白质的表达量。结果MM患者的MSCs成骨潜能和TAZ表达量较正常人降低。MM患者骨髓血清中的TNF-α浓度为（355.4±49.1）pg／ml，正常人为（92.3±17.2）pg／ml。CD138^＋骨髓瘤细胞可以抑制正常人MSCs向成骨细胞分化，TNF-α单克隆抗体可以部分逆转此抑制效应。随着培养体系中TNF-α（0、0.1、1、10ng／mL）浓度梯度增加，MSCs中TAZ表达量逐渐降低。结论MM患者的骨髓MSCs成骨潜能较正常人明显降低。骨髓瘤细胞可能是通过TNF-α抑制TAZ的表达，抑制MSCs的成骨作用。Objective To explore the relationship between TNF-α, transcriptional co-activator with PDZ-binding motif（TAZ） and bone disease of multiple myeloma. Methods The biological characteristics, especially the osteogenic potential of marrow MSCs from myeloma patients and normal subjects were studied. Real-time RT-PCR and Western-blot were employed to detect mRNA and protein expression of TAZ in MSCs. The concentration of TNF-α in the marrow plasma was detected using ELISA method. CD138^＋ myeloma cells were cocultured with normal MSCs with or without anti-human TNF-α monoclonal antibody in the Transwell system. Real-time RT-PCR was employed to detect the mRNA expressions of ALP, Chfal and TAZ in MSCs two weeks later, yon Kossa staining was used to detect the mineral deposition. TNF-α was added into the culture media of normal marrow MSCs and real-time RT-PCR and Western-blot were employed to detect mRNA and protein expression of TAZ in MSCs one week later. Results Real-time RT-PCR revealed that the mRNA of osteogenic markers was decreased in comparison with that of normal controls after cultured in the osteogenic medium, von Kossa staining showed weakened mineral deposition in MSCs from multiple myeloma patients compared with that in normal subjects after osteogenic differentiation for two weeks. The mRNA and protein levels of TAZ in the MSCs from myeloma patients were decreased. TNF-α concentration in the marrow plasma of myeloma patients was higher than that in the normal controls [（355.4±49.1） vs. （92.3±17.2） pg/ml]. CD138^＋ myeloma cells inhibited mRNA expressions of ALP, Cbfall and TAZ in MSCs, which could be partially reversed by anti-human TNF-α monoclonal antibody. Conclusion The osteogenic potential of MSCs from myeloma patients is significantly decreased in comparison with that in normal subjects, which may play an important role in the pathology of myeloma bone disease. TAZ expression inhibited by TNF-α may play an important role in this inhibition effect.

关 键 词：多发性骨髓瘤 间充质干细胞 成骨 TAZ TNF-α 

分 类 号：R686[医药卫生—骨科学]