多发性骨髓瘤患者间充质干细胞TAZ和Wnt／β-catenin基因表达及其成骨潜能的研究  

Gene expression of TAZ and Wnt/β-catenin in multiple myeloma mesenchymal stem cells and its potential osteogenesis

作　　者：王宁 黄瑞宏[2] 李祎[2] 楼瑾[2] 张琼丽[2] 李明[2] 杜新[2] 卓家才[2] 

机构地区：[1]深圳市光明新区公明人民医院血液科,518106 [2]深圳市第二人民医院深圳市血液研究所

出　　处：《白血病．淋巴瘤》2011年第11期652-655,共4页Journal of Leukemia & Lymphoma

基　　金：深圳市科技信息局科研基金（201002034）

摘　　要：目的研究多发性骨髓瘤（MM）患者骨髓间充质干细胞（MSC）成骨诱导后成骨转录共刺激因子（TAZ）和Wnt／β-cateninmRNA的表达，探讨多发性骨髓瘤骨病（MBD）潜在的治疗靶点。方法体外分离培养10例初治MM患者及10名健康对照者骨髓MSC，体外诱导分化为成骨细胞，茜素红实验检测矿化物沉积、荧光定量反转录聚合酶链反应（RT-PCR）方法检测骨桥蛋白（OPN）、骨钙素（OC）、碱性磷酸酶（ALP）、核心结合因子α1（Cbfα1）mRNA的表达，分析成骨细胞分化指标变化；荧光定量RT-PCR方法检测成骨细胞的TAZ、Writ／β-eateninmRNA，分析两组间TAZ和Wnt／β—eateninmRNA表达差异。结果骨髓MSC体外成骨诱导后，茜素红染色阳性，呈现明显红色钙化结节；实验组MSC诱导后与诱导前相比，成骨细胞分化指标OC、ALP、Cbfα1mRNA表达增高[（2．0958±0．5665、2．6670±0．3847、0．8463±0．3473）比（1．3487±0．9291、1．1452±0．6054、0．4439±0．2945）1（t值分别为2．171、6．709、2．795；均P〈0．05）；对照组MSC诱导后与诱导前相比，成骨细胞分化指标OPN、OC、ALP、Cbfα1mRNA表达增高f（2．1096±0．8267、2．8991±0．3531、4．3045±0．2844、1．3273±0．4075）比（1．2200±0．9091、0．8780±0．3927、1．9161±0．2684、0．6736±0．2513）]（t值分别为2．289、12．103、25．134、4．411；均P〈0．05）。MSC体外诱导成骨细胞，实验组与对照组相比，成骨指标OPN、OC、ALP、Cbfoa1mRNA表达降低[（1．2710±0．5636、2．0958±0．5665、2．6670±0．3847、0．8463±0．3473）比（2．1096±0．8267、2．8991±0．3531、4．3045±0．2844、1．3273±0．4075）1（t值分别为-2．650、-3．805、-10．822、-2．841；均P〈0．05）。实验组骨髓MSC体外成骨诱导后TAZ、β-eateninmRNA（2．2315±1．0723、0．5801±0．2159）比对照组（4．4140±0．8325、0．9516±0．292）表达降低（t值分别�Objective To investigate the gene expressions of TAZ and Wnt/β-catenin on the post- osteogenic cells of mesenehymal stem cells （MSC） in multiple myeloma （MM） patients and to explore the potential therapeutic target of multiple myeloma bone disease （MBD）. Methods Bone marrow mononuclear cells MNC from MM and controls were isolated, cultured, expanded and then induced to osteogenic differentiation. Realtime quantitative RT-PCR was employed to detect the osteogenic markers （TAZ, Wnt/β-catenin, OPN, OC, ALP and Cbf a1）; and alizarin red staining for mineral deposition. The mRNA expressions of TAZ and Wnt/ β-catenin in the two groups were analysed. Results Alizarin red staining was positive and the red calcium nodules were appeared on the post-osteogenic cells of MSC. The mRNA expressions of OC, ALP and Cbf ctl were 2.0958±0.5665, 2.6670±0.3847, 0.8463±0.3473, respectively, on the post-osteogenic cells of MSC in the experimental groups, which were significantly higher than those of pre-osteogenic cells （1.3487±0.9291, 1.1452±0.6054, 0.4439±0.2945） （t =2.171, 6.709, 2.795; all P 〈 0.05）. The mRNA expressions of OPN, OC,ALP and Cbf a1 were 2.1096±0.8267, 2.8991±0.3531, 4.3045±0.2844, 1.3273±0.4075, respectively, on the post-osteogenic cells of MSC in the controls, which were significantly higher than those of pre-osteogenic cells （1.2200±0.9091, 0.8780±0.3927, 1.9161±0.2684, 0.6736±0.2513） （t =2.289, 12.103, 25.134, 4.411; all P 〈 0.05）. The mRNA expressions of OPN, OC, ALP, Cbf al were 1.2710±0.5636, 2.0958±0.5665, 2,6670± 0.3847, 0.8463±0.3473, respectively, on the post-osteogenic cells of MSC in the experimental groups, which were significantly lower than those of control groups （2.1096±0.8267, 2.8991±0.3531, 4.3045±0.2844, 1.3273±0.4075） （t = -2.650, -3.805, -10.822, -2.841; all /9 〈 0.05）. The mRNA expression of TAZ and [3-eatenin were 2.2315±1.0723 and 0.5801±0.2159 on the post-osteogenic ceils of MSC in MM patients, which were significantly

关 键 词：骨髓 间质干细胞 成骨细胞 多发性骨髓瘤骨病 

分 类 号：R733.3[医药卫生—肿瘤]