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多发性骨髓瘤相关基因MMSA-1的表达及定位研究 被引量:1

The research on the expression and localization of multiple myeloma associated antigen MMSA-1
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摘要 目的:研究多发性骨髓瘤(MM)相关基因MMSA-1的表达谱,蛋白表达水平与细胞增殖的关系及其细胞定位。方法:应用实时荧光定量PCR检测MMSA-1基因在MM、白血病、非肿瘤疾病患者及正常人中的表达水平,并同DKK1基因进行相关比较。使用不同浓度伊班磷酸对骨髓瘤RP-MI8226细胞进行处理,流式细胞术(FCM)检测处理后8226细胞细胞周期的变化及早期凋亡情况,免疫组化法检测8226细胞MMSA-1蛋白表达水平的变化。构建pCMV-Myc真核表达载体,采用抗体免疫组化法确定MMSA-1蛋白的细胞定位。结果:MMSA-1基因在MM、白血病、非肿瘤疾病及正常人中均有表达,在MM细胞中mRNA表达显著高于其他组;MM-SA-1与DKK1基因在MM中的高表达具有一致性,且含量呈正相关。伊班磷酸可以使8226细胞生长受阻于S期,并减少细胞成熟促进因子的释放,从而使细胞周期停滞,促进其早期凋亡,同时可以下调MMSA-1蛋白的表达。MMSA-1主要表达于细胞膜上,但在细胞质内也可见少量蛋白。结论:MMSA-1与MM细胞增殖和溶骨破坏密切相关,为进一步研究其功能及基于该抗原的免疫治疗奠定了基础。 AIM: To study the expression profile of mul- tiple myeloma associated gene ( MMSA-1 ), explore the relationship between its expression level and MM cells' prolif- eration as well as its celluler localization. METHODS: The mRNA levels of MMSA-1 and DKKI genes were detected by RT-PCR in patients with MM, leukemia, non-tumor diseases and in the healthy donors, respectively. Then, their correlation was analyzed. The effects of Ibandronate Sodi- um on the cell cycle and early apoptosis of 8226 cells were analyzed by flow cytometry, and the effect on its protein expression was analyzed by immunohistochemistry. Construct MMSA-I eukaryotic expression vector pCMV-Myc-MMSA-I, and antibody immunohistochemistry was applied to study the cellular localization of the protein. RESULTS: MMSA-I gene was expressed in all of the specimens described above, and the mRNA level in MM was much higher than that in the others, just like DKK1 gene. More than that, their expression exhibited a significant positive correlation. Ibandronate Sodium could inhibit cell proliferation by a cell-cycle arrest in Sphase. By reducing cell maturation promoting factor release, it stopped the cell cycle, promoted their early apoptosis and decreased the protein expression of MMSA-I. MM- SA-I protein principally distributed on cell membranes, however, there are a small quantity in cytalplasm. CONCLU- SION: These results revealed that MMSA-I may play a pivotal role in MM proliferation and osteolysis destruction, which lay the foundation for the further study of biological function and immunotherapy based on MMSA-I.
出处 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2012年第1期63-66,71,共5页 Chinese Journal of Cellular and Molecular Immunology
基金 国家自然科学基金青年基金(30700337) 西安市科技计划项目资助(HM1117(4))
关键词 多发性骨髓瘤 MMSA-1 DKK1 细胞定位 multiple myeloma MMSA-1 DKK1 cellularlocalization
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