口腔医学研究

口腔医学研究 ›› 2017, Vol. 33 ›› Issue (5): 525-529.DOI: 10.13701/j.cnki.kqyxyj.2017.05.015

• 临床研究论著 • 上一篇    下一篇

LMP-1转染人胎盘源间充质干细胞前后蛋白质组学研究

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   

  1. 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
  • 收稿日期:2016-12-02 出版日期:2017-05-20 发布日期:2017-05-26
  • 通讯作者: 刘志辉,E-mail:liuzhihui1975@sina.com
    王博蔚,E-mail:wanbowei2009@hotmail.com
  • 作者简介:宋立杰(1990~ ),女,河北沧州人,硕士研究生,主要研究方向:口腔颌面部组织工程骨研究。
  • 基金资助:
    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)

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

SONG Li-jie1, YANG Jun-xing1, WANG Yao1, LU Ya-dong1, YANG Nan1, JIN Xin1, SUN Meng-jie1, LIU Zhi-hui1*, WANG Bo-wei2*, ZHU Zhen1.   

  1. 1. Department of Prosthodontics, Jilin University, Changchun 130021, China; 2. Department of Obstetrics and Gynecology, The Second Hospital of Jilin University, Changchun 130021, China.
  • Received:2016-12-02 Online:2017-05-20 Published:2017-05-26

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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。

关键词: LMP-1, hPDMSCs, 蛋白质组学

Abstract: 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-1 and 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, eEF2 and peroxiredoxin-1) which were closely related to osteogenesis and had a higher MALDI-TOF-MS-MS



基金项目 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)
作者简介 宋立杰(1990~ ),女,河北沧州人,硕士研究生,主要研究方向:口腔颌面部组织工程骨研究。
*通讯作者 刘志辉,E-mail:liuzhihui1975@sina.com
王博蔚,E-mail:wanbowei2009@hotmail.com

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 hPDMSCsLMP-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, 9 proteins 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-1 on hPDMSCs cell function and osteogenic differentiation.

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