肺癌组织蛋白质组双向凝胶电泳方法的建立和优化

目的建立和优化肺癌组织蛋白质组双向凝胶电泳(2-DE)技术,获取高分辨率、重复性好的蛋白质2-DE图谱。方法提取肺癌组织蛋白,以固相pH梯度胶条做第一向等电聚焦电泳,十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)为第二向,在上述过程中分别对蛋白提取、等电聚焦电泳和凝胶染色进行控制和优化,利用ImageMaster 2Dplatinum 6.0分析软件获得二维凝胶图像。结果实验重复3次,共获15幅二维凝胶图像,平均蛋白质点数为1 137±57;随机选取1例标本的2幅图像,利用软件对2幅图像中蛋白点进行匹配,匹配率为90.4%。结论优化肺癌组织蛋白质组2-DE技术可获得高分辨率、重复性好的蛋白质2-DE图谱,为开展肺癌组织蛋白质组学研究打下基础。

Establishment and optimization of two-dimensional polyacrylamide gel electrophoresis for proteom analysis of lung tumor tissue

Objective To establish and optimize the technique of two-dimensional polyacrylamide gel electrophoresis(2-DE) for proteom analysis of lung tumor tissue in order to obtain 2-DE protein map with high-resolution and better repeatability.Methods Proteins from lung tumor tissue were extracted.Immobilized pH gradient(IPG) strips was used to perform isoelectric focusing electrophoresis(the first dimension),and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) served as the second dimension.In procedures above,protein extraction,isoelectric focusing electrophoresis and gel staining were controlled and optimized,respectively.ImageMaster 2D platinum 6.0 analyzing software was used to gain two-dimensional gel images.Results The experiment was repeated three times,and a total of 15 two-dimensional gel images were obtained with average protein spots of 1 137±57.Two images of a sample were selected randomly.Matching the protein spots in the 2 images using software demonstrated a matching rate of 90.4%.Conclusion 2-DE protein map with high-resolusion and better repeatability can be obtained by optimizing 2-DE technique for proteom analysis of lung tumor tissue,and lay the foundation for further proteomics research of lung tumor.