论文部分内容阅读
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 a method for in vitro enrichment of protein secreted by colonic carcinoma in mice with colitis. METHODS: Colon of mice with colitis-associated colon cancer was used in this study. Colon of mice in different stages were cultured in vitro for 48h to collect secretory proteins. Secreted proteins were separated and analyzed by SDS-PAGE, and the presence or absence of cytoplasmic protein or nuclear protein contamination of the secreted protein was detected by Western blot. Results: The secreted proteins collected under the experimental conditions had good reproducibility, less contamination of nucleoprotein and cytoplasmic protein in the secretory proteins, and could distinctly distinguish the protein bands differentially expressed in the course of the disease. Conclusion: The in vitro enrichment of colonic-associated colon cancer secreting protein has been successfully established.