| 摘要: |
| 目的:建立RASSF1A表达或表达缺失的两类不同早期肺腺癌组织蛋白质的双向电泳凝胶图谱，筛选并鉴定存在明显表达差异的蛋白质。方法:采用Western印迹技术,从RASSF1A转录缺失和RASSF1A转录正常的早期肺腺癌标本中筛选出RASSF1A表达和表达沉默的癌组织各5例。提取组织可溶性总蛋白，采用固相pH梯度双向凝胶电泳技术分离总蛋白，建立两类不同早期肺腺癌组织蛋白质的双向电泳凝胶图谱；PDQuest凝胶图像分析软件比较分析，筛选出差异表达的蛋白质点；基质辅助激光解吸电离飞行时间质谱获得相应蛋白质点的肽质量指纹图谱；搜索蛋白质数据库鉴定差异表达的蛋白质。结果:建立了重复性较好的RASSF1A表达或表达缺失的两类不同早期肺腺癌组织蛋白质双向电泳凝胶图谱；筛选出存在明显表达差异的蛋白质点17个，挖取其中的9个进行质谱分析，9个蛋白质点均得到了满意的肽质量指纹图谱；搜索蛋白质数据库鉴定出5种蛋白质，分别是：细胞色素b5（cytochrome b5），60S磷酸核糖体蛋白P2（60S acidic ribosomal protein P2），碳酸酐酶1（carbonic anhydrase1），5吡咯啉羧酸还原酶1（pyrroline5carboxylate reductase1）和载脂蛋白AⅠ前体蛋白（apolipoprotein AⅠ precursor）。结论:成功建立了RASSF1A表达或表达缺失的两类不同早期肺腺癌组织蛋白质的双向电泳凝胶图谱，从中鉴定出5种存在明显表达差异的蛋白质，为进一步研究RASSF1A作用的信号转导途径奠定了初步的基础。 |
| 关键词: 肺肿瘤 腺癌 RASSF1蛋白 蛋白质组学 双向凝胶电泳 基质辅助激光解吸电离飞行时间质谱 |
| DOI：10.3724/SP.J.1008.2008.00136 |
| 投稿时间：2007-10-31修订日期：2007-12-11 |
| 基金项目:国家自然科学基金（30571552）. |
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| Screen and identification of differential proteins in two early stage lung adenocarcinoma tissues with and without RASSF1A expression |
| LIU Gui-zhi1,WU Yi-ming2* |
| (1. Department of Physical Examination, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China;2. Department of Occupational Health and Toxicology, College of Public Health, Zhengzhou University, Zhengzhou 450052) |
| Abstract: |
| Objective：To establish a twodimensional electrophoresis (2DE) gel map of 2 earlystage lung adenocarcinoma tissues with and without RASSF1A expression, so as to screen and identify differential proteins. Methods: Five earlystage lung adenocarcinoma tissues with RASSF1A expression and 5 without RASSF1A expression were screened out by Western blotting assay. The total soluble proteins of the tissue were extracted and were separated by immobilized pH gradient based twodimensional gel electrophoresis to set up the 2DE gel map of the 2 adenocarcinoma tissues. The differentiallyexpressed proteins were analyzed by PDQuest image analysis software and identified by matrixassisted laser desorption/ionization time of flight mass spectrometry (MALDITOFMS); the protein database was searched to further characterized the differential proteins. Results: A wellreproducible 2DE gel map of the 2 adenocarcinoma tissues with and without RASSF1A expression was established and 17 differential protein spots were screened out. Nine of 17 differential protein spots were selected for MALDITOFMS study and satisfactory peptide mass fingerprints were obtained for all the 9 spots. Searching of the protein database revealed 5 candidate proteins and they were: cytochrome b5, 60S acidic ribosomal protein P2, carbonic anhydrase 1, pyrroline5carboxylate reductase 1, and apolipoprotein AⅠ precursor.Conclusion: We have successfully obtained the 2DE gel images of 2 earlystage lung adenocarcinoma tissues with and without RASSF1A expression, and from which we have identified 5 differential proteins,which paves a way for studying the signal transduction pathways involving RASSF1A. |
| Key words: lung neoplasms adenocarcinoma RASSF1 protein proteomics two dimensional gel electrophoresis matrix assisted laser desorption ionization time of flight mass spectrometry |
