摘要
目的 分析吲哚美辛(indomethacin,IN)对人结肠癌细胞系HCT116蛋白质表达谱的影响,为研究IN抗结肠癌作用的相关蛋白提供新的方法。方法 应用固相pH梯度(immobilizedpHgradient,IPG)双向凝胶电泳技术(two dimensionalgelelectro phoresis,2 DE)分离IN 处理组和未处理组HCT116细胞的总蛋白,银染显色,ImageMaster2D图像分析软件比较分析IN 处理组和未处理组HCT116细胞系的双向凝胶电泳图谱并识别差异表达的蛋白质。结果 获得了背景清晰、分辨率高、重复性好的HCT116细胞系的双向凝胶电泳图谱, IN 未处理组与处理组蛋白质点数为:1256±50,1169±36;匹配率为:93.0%,90.6%。IN 未处理组的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等电聚焦(isoelectricfocusing,IEF)方向的偏差是0.896±0.177mm,在十二烷基磺酸钠聚丙烯酰胺凝胶电泳(sodiumdo decylsufate polyacrylamidegelelectrophoresis,SDS PAGE)方向的偏差为1.106±0.289mm。比较分析IN 处理组和未处理组HCT116细胞的双向凝胶电泳图谱,发现IN 处理组和未处理组共有45个差异点,IN 处理组有9个蛋白质点表达上调,34个蛋白质点表达下调,2个蛋白质点仅在IN 未处理组表达。结论 首次建立了IN 处理和未处理HCT116细胞的双向凝胶电泳图谱,识别?
Objective To study the variation of protein maps for indomethacin (IN)-treated human colon cancer cell line (HCT116) using two-dimensional Gel Electrophoresis (2-DE) and ImageMaster analysis. Methods Proteome profiles of HCT116 were established in IN-treated and untreated groups,which separated by immobilized pH gradient-based two-dimensional gel electrophoresis(2-DE). The gels were stained by silver, scanned by ImageScanner, analyzed with ImageMaster software. Results Clear background, well-resolved and reproducible 2-DE patterns of HCT116 cells were acquired in IN-treated and untreated groups. 1256±50, 1169±36 protein spots were resolved respectively in IN-untreated and IN-treated group. The match rate in IN- untreated group was 93.0%, It was 90.6% in IN-treated group. The average deviations of spot position were 0.896±0.177mm in IEF direction and 1.106±0.289mm in SDS-PAGE direction, respectively. Forty five differential protein spots were displayed between IN-treated and untreated groups. Of which, 34 decreased in abundance and 9 showed higher expression in IN-treated group, 2 only expressed in IN-untreated cells. Conclusion 2-DE profiles of IN-treated and of IN-untreated HCT116 cells were established. Apparent proteome difference is detected between IN-treated and untreated HCT116 cells. Further analysis on the differential protein spots may serve as a new way to study the molecular mechanism of IN on treating Colon cancer.
出处
《肿瘤》
CAS
CSCD
北大核心
2005年第1期28-32,共5页
Tumor
基金
国家自然科学基金资助项目(编号:30271516)
