LMP-1转染人胎盘源间充质干细胞前后蛋白质组学研究  

Comparative Proteomics Analysis of hPDMSCs before and after Rransfection with LMP-1

作　　者：宋立杰[1] 杨军星[1] 王瑶[1] 卢亚东[1] 杨楠[1] 金鑫[1] 孙梦洁[1] 刘志辉[1] 王博蔚[2] 朱镇[1] 

机构地区：[1]吉林大学口腔医院,吉林长春130021 [2]吉林大学第二医院,吉林长春130022

出　　处：《口腔医学研究》2017年第5期525-529,共5页Journal of Oral Science Research

基　　金：吉林省医药产业推进计划(编号:20140311088YY;201603028YY);长春市重大科技攻关计划(编号:2014054);吉林省科技攻关计划(编号:20150204004YY);长春市"双十工程"计划(统带:16SS12)

摘　　要：目的:采用蛋白质组学技术检测LIM矿化蛋白-1(LIM mineralization protein-1,LMP-1)转染人胎盘源间充质干细胞(Human placenta-derived mesenchymal stem cells,hPDMSCs)前后蛋白质组学改变。方法:提取hPDMSCs LMP-1和hPDMSCsvector细胞总蛋白,通过二维凝胶电泳(two-dimensional gel electrophoresis,2-DE)制备两组细胞的二维电泳图谱,并用PDQuest软件进行图像分析,寻找差异表达的蛋白质点,并用基质辅助激光解吸粒子-飞行时间电离质谱(matrix-assisted laser desorption-ionization time-of-flight tandem mass spectrometry,MALDI-TOF-MS)分析技术对这些差异表达的蛋白质点进行鉴定,并选取6个与成骨相关的蛋白(PCNA,vimentin,annexin A1,annexin A2,eEF2和peroxiredoxin-1)作为验证对象,蛋白质印迹法检测各蛋白在两组细胞中的表达水平。结果:采用2-DE技术建立了hPDMSCsLMP-1和hPDMSCsvector两组细胞的二维电泳图谱;PDQuest软件分析图像发现两组细胞有22个差异蛋白质点,采用MALDI-TOF-MS质谱初步鉴定出15个蛋白质点,其中表达上调9个,下调6个。Western blot检测结果与蛋白质组学一致。结论:LMP-1基因转染hPDMSCs后共筛选获得15个差异表达的蛋白,它们是成骨分化潜在的重要参与者,为进一步阐明LMP-1对hPDMSCs细胞功能和成骨分化影响的分子机制奠定基础。Objective：To explore the protein component changes in human placenta-derived mesenchymal stem cells（hPDMSCs）transfected with LIM mineralization protein-1（LMP-1）gene using proteomics technologies.Methods：The hPDMSCsLMP-1and hPDMSCsvector were collected to extract total proteins.The two-dimensional gel electrophoresis（2-DE）of total proteins in cells of two groups was created.The 2-DE map was analyzed and compared using PDQuest software to find out differentially expressed protein spots.These differentially expressed candidate proteins were identified by matrix-assisted laser desorption-ionization time-of-flight tandem mass spectrometry（MALDI-TOF-MS）.Six protein spots（PCNA,vimentin,annexin A1,annexin A2,eEF2and peroxiredoxin-1）which were closely related to osteogenesis and had a higher MALDI-TOF-MS-MS score were selected and detected by Western blotting.Results：The 2-DE maps of total proteins in cells were successfully established.Twenty-two differentially expressed protein spots with a difference in expression between hPDM-SCsLMP-1 and hPDMSCsvector were identified by PDQuest 2-D Analysis Software.Combination of MALDI-TOF-MS and database search identified a total of 15 differentially expressed protein spots.Of all the identified protein spots,9proteins were up-regulated and 6 proteins were down-regulated in hPDMSCsLMP-1 as compared with the hPDMSCsvector.Western blot results were fully consistent with proteomics.Conclusion：Fifteen differentially expressed proteins in hPDMSCsLMP-1 were identified.They were potentially important participants in osteogenic differentiation,laid the foundation for further molecular mechanism elucidation of LMP-1on hPDMSCs cell function and osteogenic differentiation.

关 键 词：LMP-1 hPDMSCs 蛋白质组学 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]