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-associated Colon Cancer Secretome

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。

Objective： To establish in vitro method to collect the secreted proteins from mouse with colitis-associated colon can- cer. 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 semm-fi＇ee 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 pro- teins from mouse with colitis-associated colon cancer was successfully established.