论文部分内容阅读
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。
OBJECTIVE: To investigate the expression of MMSA-1 in multiple myeloma (MM) and its relationship with cell proliferation and its cellular localization. Methods: The expression of MMSA-1 gene in patients with MM, leukemia, non-neoplastic disease and normal controls was detected by real-time fluorescent quantitative PCR and compared with DKK1 gene. The myeloma RP-MI8226 cells were treated with different concentrations of ibandronic acid and the cell cycle and early apoptosis of 8226 cells were detected by flow cytometry (FCM). The expression of MMSA-1 protein in 8226 cells was detected by immunohistochemistry Horizontal changes. The eukaryotic expression vector pCMV-Myc was constructed and the cellular localization of MMSA-1 protein was determined by immunohistochemistry. Results: MMSA-1 gene was expressed in MM, leukemia, non-neoplastic disease and normal human. The mRNA expression of MMSA-1 in MM was significantly higher than that in other groups. The high expression of MM-SA-1 and DKK1 in MM was consistent Sex, and the content was positively correlated. Ibandronic acid inhibited the growth of 8226 cells in S phase and decreased the release of cell maturation promoting factors, which then arrested cell cycle and promoted its early apoptosis, meanwhile down-regulated the expression of MMSA-1 protein. MMSA-1 is mainly expressed on the cell membrane, but also a small amount of protein can be seen in the cytoplasm. Conclusion: MMSA-1 is closely related to the proliferation of MM cells and the destruction of osteolytic bone, which lays the foundation for further study of its function and immunotherapy based on this antigen.