人肺鳞癌；肺炎性假瘤；固相pH梯度双向凝胶电泳；MALDI

人肺鳞癌及肺炎性假瘤组织的蛋白质差异表达谱的初步建立

http://www.100md.com 《中华现代内科学杂志》 2006年第8期

人肺鳞癌；肺炎性假瘤；固相pH梯度双向凝胶电泳；MALDITOFMS；,蛋白质组,,],人肺鳞癌；肺炎性假瘤；固相pH梯度双向凝胶电泳；MALDITOFMS；,蛋白质组,人肺鳞癌及肺炎性

     [摘要] 目的建立分辨率高和重复性好的人肺鳞癌组织及其肺炎性假瘤组织的双向凝胶电泳图谱，并识别鉴定出其差异表达的蛋白质。方法利用固相pH梯度双向凝胶电泳分离人肺鳞癌组织及肺炎性假瘤组织的总蛋白质，凝胶经银染显色后，运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质，应用基质辅助激光解吸电离飞行时间质谱(MALDITOFMS)及数据库搜索鉴定部分差异蛋白质点。结果 (1)得到了分辨率较高、重复性较好的人肺鳞癌组织和肺炎性假瘤组织的双向凝胶电泳图谱，将同一癌组织和肺炎性假瘤组织分别进行了3次双向凝胶电泳，3块凝胶的平均蛋白质点数分别为1569±78和1487±64，平均匹配的点数分别为1454±37和1370±43，匹配率达92.7%和92.1%；同一癌组织的3块胶在蛋白质点位置上有较好的重复性，不同胶间蛋白质点在IEF方向的偏差是(0.733±0.101)mm，在SDSPAGE方向上的偏差为(0.925±0.207)mm。(2)比较分析肺鳞癌组织及肺炎性假瘤组织的双向凝胶电泳图谱，平均匹配的点数为1142±51，对23个差异蛋白质点进行了肽质指纹图分析，鉴定出一些与细胞周期调控、信号传导等有关的蛋白质。结论建立了分辨率较高且重复性较好的人肺鳞癌组织与肺炎性假瘤组织的双向凝胶电泳图谱,并识别鉴定出了一些差异表达的蛋白质，这不仅有助于进一步完善人肺癌蛋白质组数据库，而且能为人肺鳞癌发病机制研究及其诊断、鉴别诊断、治疗以及新药开发等提供线索。

    [关键词] 人肺鳞癌；肺炎性假瘤；固相pH梯度双向凝胶电泳；MALDITOFMS；蛋白质组

    Establishing and analyzing differential proteomic expression profiles from human lung squamous carcinoma and inflammatory pseudotumor tissues

    LI Cui,DUAN Chaojun,XIAO Zhiqiang,et al.

    Key Laboratory of Cancer Proteomics of Ministry of Health,Xiangya Hospital,Central South University,Changsha 410008,China

    [Abstract] Objective To establish differential proteomic expression analysis of high resolution and reproducibility twodimensional electrophoresis profiles from human lung squamous carcinoma tissues and inflammatory pseudotumor tissues.Methods The proteome of human lung squamous carcinoma tissues and inflammatory pseudotumor tissues were separated and identified by using a series of methods,including immobilized pH gradient twodimensional polyacrylamide gel electrophoresis,silver staining,ImageMaster 2DE analysis software,peptide mass fingerprint based on matrixassisted laser desorption/ionization time of flight mass spectrometry (MALDITOFMS) and database searching.Results (1) The good 2DE pattern of human lung squamous carcinoma tissues and inflammatory pseudotumor tissue including resolution and reproducibility were obtained.After staining,the 2DE image analysis by ImageMaster 2DE software had detected average of 1569±78 and 1487±64 spots,1454±37 and 1370±43 spots were matched individually.The average matching rate were 92.7% and 92.1%.There had a good reproducibility of spot position in the human lung squamous carcinoma tissues maps,with average deviation in IEF direction of (0.733±0.101)mm,while in SDSPAGE direction it was (0.925±0.207)mm.(2) The map of the tumor tissues was compared with inflammatory pseudotumor tissues,1142±51 spots were matched.Twentythree proteins were analyzed,some of which have the relationship of oncogenes,and others were the regulation proteins,which adjust the cell cycle and signal transduction.Conclusion These data suggest that we constructed high resolution and reproducibility twodimensional electrophoresis profiles from human lung squamous carcinoma tissues and inflammatory pseudotumor tissues and analyze the differential protein spots,which will be taken advantage to establishing the differential proteomic expression profiles.It is suggested that the differential expression analysis of proteome may be useful to further study of human lung squamous carcinoma. ......

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