乳腺增生组织蛋白质组的凝胶内差异显示电泳分析

Proteome study of hyperplasia of mammary glands by two-dimensional difference gel electrophoresis

目的：建立具有高分辨率和稳定性的乳腺增生组织蛋白质组的双向电泳图谱,并对其进行差异蛋白质组分析。方法：取乳腺增生病患者增生部位及正常部位乳腺组织,匀浆提取乳腺组织总蛋白,分别用Cy3或Cy5标记,每一对Cy3和Cy5标记样品都与一个Cy2标记的内标等量混合,上样于同一胶中进行电泳分离,经不同光激发下扫描得到不同样品的蛋白质组图谱。所获得的图谱经DeCyder软件进行分析。结果：在乳腺增生病增生的组织中,有12个蛋白质表达水平显著增加,另外3个蛋白质表达水平显著下降。结论：利用DIGE技术可以作胶内对比分析,也可以根据内标消除胶与胶之间的差异,提高统计的可信度；分析所得的15个差异蛋白质可能与乳腺增生疾病的发生与发展有关。

Objective： To establish and analysis maps of two - dimensional difference gel electrophoresis for proteomic study of hyperplasia of mammary glands. Methods： Normal mammary gland.s and surgically reseeted hyperplasia of mananary glands constitution were homogenized with lysis solution. The extracts were labeled with Cy3 or Cy5 randoudy, Each Cy3 - labeled sample and Cy5 - labeled sample were mixed on the santa 2 - D gel along with a Cy2 - labeled mixture of all samples as an internal standard. The gels were respectively imaged by Typhoon 9400 using different emission filter and spot - features were analyzed by DeCyder software in each paired normal/disease comparison. Result： 12 unique proteins that were changing in abundance more than 1,4 folder up wiile 3 proteins decreased more titan i .4 folder down between normal mammary glands and surgically resected hyperplasia of mammary glands constitution. Conclusions： using DIGE technology with internal standard, statistically significant quantitative comparisons of each protein abundance change could be made across multiple samples simultaneously without interference due to gel - to - gel variation, And these 15 proteins might be involved in the process of hyperplasia of mammary glands.