LMP-1转染人胎盘源间充质干细胞前后蛋白质组学研究

发布时间：2018-03-20 17:27

本文选题：LMP-　切入点：hPDMSCs　出处：《口腔医学研究》2017年05期 　论文类型：期刊论文

【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。
[Abstract]:Objective: using proteomics technique for the detection of LIM (LIM mineralization protein-1 -1 mineralization protein, LMP-1) transfection of human placenta derived mesenchymal stem cells (Human placenta-derived mesenchymal stem cells, hPDMSCs) before and after the proteome changes. Methods: the extraction of hPDMSCs LMP-1 and hPDMSCsvector cell total proteins by two-dimensional gel electrophoresis (two-dimensional gel electrophoresis, 2-DE two dimensional electrophoresis) preparation of cells in two groups, and the images were analyzed by PDQuest software to find the differentially expressed proteins, and by matrix assisted laser desorption time-of-flight particle ionization mass spectrometry (matrix-assisted laser desorption-ionization time-of-flight tandem mass spectrometry, MALDI-TOF-MS) analysis of these differentially expressed proteins were identified, and selected 6 a protein and bone related (PCNA, vimentin, annexin A1, annexin A2, and eEF2 Peroxiredoxin-1) as a validation object, the expression level of each protein by Western blotting in two groups of cells. Results: two dimensional electrophoresis of hPDMSCsLMP-1 and hPDMSCsvector of two groups were established by 2-DE technology; image found in the two groups of cells have 22 differential protein analysis software PDQuest, using MALDI-TOF-MS mass spectrometry identified 15 protein spots among them, 9 up-regulated, 6 down regulated.Western blot detection results and proteomics. Conclusion: the transfection of LMP-1 gene after hPDMSCs were screened 15 differentially expressed proteins obtained, they are an important participant in osteogenic differentiation potential, in order to further clarify the molecular mechanism of LMP-1 on hPDMSCs cell function and osteogenic differentiation of lay the foundation.

【作者单位】：吉林大学口腔医院;吉林大学第二医院;
【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)
【分类号】：R329.2

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