色谱

色谱

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用于相对和绝对定量的等量异位标签-二维液相色谱-串联质谱法分析雌二醇和血清剥夺对乳腺癌细胞内蛋白质含量的影响

朱镭1, 倪国新1, 张政希1, 徐学敏1,2, 胡晓芳2, 李伟1*   

  1. 1.Institute for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, China; 2.School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
  • 收稿日期:2008-09-22 修回日期:2008-11-24 出版日期:2009-05-30 发布日期:1982-09-25
  • 通讯作者: 李伟

Analysis of the effects of 17β-oestradiol and serum deprivation on the contents of proteins in breast cancer cells by isobaric tags for relative and absolute quantification and two-dimensional liquid chromatography-tandem mass spectrometry

ZHU Lei1, NI Guoxin1, ZHANG Zhengxi1, XU Xuemin1,2, HU Xiaofang2, LI Wei1*   

  1. 1.Institute for Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, China; 2.School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
  • Received:2008-09-22 Revised:2008-11-24 Online:2009-05-30 Published:1982-09-25
  • Contact: LI Wei

PDF

795

被引次数

13

摘要： 雌激素和血清中的激素及各种生长因子在乳腺癌的发生、发展过程中发挥着重要的作用。研究雌激素和血清对乳腺癌细胞中蛋白质组成和含量的影响对于阐明雌激素和血清对乳腺癌细胞影响的分子机理具有重要的意义。利用四重用于相对和绝对定量的等量异位标签(isobaric tags for relative and absolute quantification, iTRAQ)标记结合二维液相色谱-串联质谱(two-dimensional liquid chromatography-tandem mass spectrometry, 2D-LC-MS/MS)对雌二醇(17β-oestradiol, E2)和血清各自以及共同作用对MCF7乳腺癌细胞内蛋白质表达的影响进行了比较,共鉴定到置信度在95%以上的蛋白质576种,其中各组相对于正常培养组的细胞共找到26种差异在1倍以上的蛋白质,其中10种上调,16种下调。研究发现E2和血清可显著影响细胞内与蛋白质合成相关的蛋白质的水平。实验结果表明: iTRAQ-LC-MS/MS是进行多个样品差异蛋白组比较的一种有效方法。

关键词: 雌激素, 二维液相色谱-串联质谱, 乳腺癌细胞 , 血清剥夺, 用于相对和绝对定量的等量异位标签(iTRAQ)

Abstract: Estrogen, hormones and growth factors in serum play important roles in the carcinogenesis and development of breast cancer. The study of the effects of the above factors on protein components and their contents in breast cancer cells is necessary for understanding the molecular mechanisms. In this study, isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled with two-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS) was employed to compare the differential protein expressions in MCF7 cells upon 17β-oestradiol (E2) stimulation or serum deprivation (SD) versus in normal conditions. A total of 576 proteins with the confidential level of ≥95% were identified, 26 proteins among the 576 proteins were found to be differentially expressed after E2 stimulation or serum deprivation, and among them 10 proteins were up-regulated and 16 proteins were down-regulated by more than two folds. Interestingly, both E2 and serum significantly altered the level of proteins involved in protein synthesis. This demonstrated iTRAQ coupled LC-MS/MS is an efficient methodology in comparative proteomics.

Key words: breast cancer cells , estrogen, serum deprivation, two-dimensional liquid chromatography-tandem mass spectrometry (2D-LC-MS/MS), isobaric tags for relative and absolute quantification (iTRAQ)