初发和复发/难治多发性骨髓瘤细胞蛋白谱表达差异  

Differential Protein Expression in Primary and Relapsed or Refractory Multiple Myeloma Cells

作　　者：冯慧 刘念[2] 刘健[2] 陈文明[2] FENG Hui;LIU Nian;LIU Jian;CHEN Wenming(Department of Hematology, Beiijing Pinggu Hospital,Capital Medical University, Beijing, 101200,China;Department of Hematology, Beiijing Chaoyang Hospital, Capital Medical University, Beijing, 100020,China)

机构地区：[1]首都医科大学附属北京平谷医院血液科,北京市101299 [2]首都医科大学附属北京朝阳医院血液科,北京市100020

出　　处：《医学分子生物学杂志》2019年第4期390-394,共5页Journal of Medical Molecular Biology

基　　金：平谷区医院科学技术基金(No.pgyy2015-21号).

摘　　要：目的研究初发和复发/难治多发性骨髓瘤细胞(multiple myeloma,MM)的蛋白表达差异,寻找促进多发性骨髓瘤复发/难治的关键蛋白。方法选取初发和复发/难治多发性骨髓瘤CD138阳性原代细胞各10例为研究对象。应用蛋白质谱法分析两组细胞的表达差异蛋白。Sequest软件鉴定筛选的差异蛋白。选取筛选的差异蛋白,应用在线数据库PANTHER(http://www.pantherdb.org/)对差异蛋白进行分类,应用人生物信息数据库STRING 9.05(http://string-db.org/)分析差异蛋白之间的相互作用,筛选对细胞生物学特性具有重要影响的蛋白质。在差异蛋白中选取与MM发生、发展相关的蛋白,并结合文献报道共选出3种蛋白,对7例初发和8例复发/难治MMCD138阳性细胞,通过Western印迹的方法验证这3种蛋白的表达。结果①差异蛋白筛选:在10例初发和10例复发/难治MM原代细胞中共鉴定出1716种蛋白质,对两组细胞的蛋白表达进行比较,230种蛋白的表达差异超过1.5倍,其中63种蛋白表达下调,167种蛋白表达上调。差异蛋白的分子功能主要有催化活性、结合活性、结构分子活性、转运蛋白活性、受体活性、翻译调控活性、酶调节活性和抗氧化活性等。上调蛋白具有催化活性和结合活性的蛋白质所占比例64.4%,PANTHER、STRING分析蛋白功能及蛋白相互作用,其中8个与恶性疾病及MM进展相关的蛋白,而下调蛋白中未发现与肿瘤及多发性骨髓瘤进展相关蛋白。②差异表达蛋白验证:筛选序列覆盖率最高的Vimentin及在MM发生、发展中具有重要影响的蛋白质,Ezrin、HSP90进行验证。Western印迹实验结果证实Vimentin、Ezrin在复发/难治细胞中表达升高,有统计学差异,而HSP90在两者均有表达,但无统计学差异。结论初发和复发/难治性CD138阳性骨髓瘤细胞具有不同的蛋白表达谱,其中Ezrin、Vimentin可能与多发性骨髓瘤复发/难治有关。Objective By examining differences in the protein expression among primary and relapsed/refractory multiple myeloma (MM) cells, the study tried to find the key proteins in the relapsed or refractory cells. Methods CD138-positive cells were collected from the bone marrow of 10 cases primary MM and 10 cases of relapsed/refractory MM , and were detected for protein expres-sion by mass spectrometry (HPLC). The differential expression proteins were identified by using Sequest software package. The differentially expressed proteins were classified by the PANTHER (http://www. pantherdb. org/) and analyzed for their interactions by STRING 9. 05 ( http://string-db.org/). The proteins that exert important effect on cellular biological properties were screened. Three differential proteins related to the development and progression of MM were found. The protein expressions were verified by Western blotting. Results ① Screening of differential proteins: A total of 1716 proteins were identified in the CD 138-positive cells collected from the bone marrow of 10 cases of primary MM and 10 cases of relapsed/refractory MM. The expression differences of 230 proteins were more than 1. 5 times. Among them, the expression of 63 proteins was down-regulated and expression of 167 proteins were up-regulated. The functions of these differentially expressed proteins were mainly of catalytic, binding, structural and molecular, transportational nature or receptor-, translation regulation-enzyme activity-and anti -oxidation-related. The ratio between proteins with catalytic activity and those with binding activity was 64. 4 %. PANTHER and STRING analysis showed that 8 proteins were associated with malignant diseases and MM progression , whereas those whose expression was downregulated were not associated with malignant disease or MM progression.② Verification of the differential expressed protein: The expression of Vimentin , which has the highest sequence coverage rate and, HSP90 and Ezrin, which have important impact with the development

关 键 词：多发性骨髓瘤 CD138阳性细胞 蛋白表达谱 

分 类 号：R394.34[医药卫生—医学遗传学]