文章摘要
马怡茗肖汀赵新华韩文晓汪红英△.结肠炎相关结肠癌分泌蛋白质体外富集方法的建立*[J].,2014,14(17):3259-3261
结肠炎相关结肠癌分泌蛋白质体外富集方法的建立*
Establishment of in Vitro Methods for Collection of Colitis-associatedColon Cancer Secretome*
  
DOI:
中文关键词: 结肠炎相关结肠癌  分泌蛋白质  组织培养
英文关键词: Colitis-associated colon cancer  Secreted protein  Tissue culture
基金项目:国家重点基础研究发展规划项目(2011CB910700);国家自然科学基金重大研究计划培育项目(91129717)
作者单位
马怡茗肖汀赵新华韩文晓汪红英△ 北京协和医学院中国医学科学院肿瘤医院肿瘤研究所分子肿瘤学国家重点实验室北京100021 
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中文摘要:
      摘要目的:建立小鼠结肠炎相关结肠癌分泌蛋白质体外富集的方法;方法:利用结肠炎相关结肠癌小鼠模型,取不同阶段实验小 鼠的结肠体外培养48h,收集分泌蛋白质。使用SDS-PAGE 分离分析分泌蛋白质,并通过Western blot 检测分泌蛋白质是否存在 胞浆蛋白或者核蛋白的污染。结果:在实验条件下收集得到的分泌蛋白重现性好,分泌蛋白中核蛋白和胞浆蛋白的污染较少,且 能够明显分辨与病程相关差异表达蛋白质条带。结论:成功建立了结肠炎相关结肠癌分泌蛋白质的体外富集方法。
英文摘要:
      ABSTRACT Objective:To establish in vitro method to collect the secreted proteins from mouse with colitis-associated colon cancer. Methods:Colitis-associated colon cancer was induced by azoxymethane (AOM) and dextran sulfate sodium (DSS) in C57BL/6 mice. Colon tissue was collected at different stages during the development of colon cancer. The whole colons were sectioned into pieces for in vitro culture with serum-free media, and then the conditioned media were collected at 48h. Western blot was applied to detect whether there is cytoplasm and nucleus protein contaminant in secreted proteins. Results:The method had good reproducibility. There was trace amount of contaminants from cytoplasm and nucleus in collected secretome. And several different protein bands were observed among different groups and correlated with the stage of colitis-associated colon cancer. Conclusion:An in vitro method to collect secreted proteins frommouse with colitis-associated colon cancer was successfully established.
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