胃腺癌患者粪便蛋白质的提取及初步质谱分析  

Protein Extraction and Preliminary Research on Proteins in Stools from Patients with Gastric Adenocarcinoma

作　　者：傅淑宏[1] 魏开华[2] 李晓霞[1] 张拓[2] 龚美亮[1] 周玉[1] 李燕燕[1] 白洁[1] 

机构地区：[1]解放军总医院南楼检验科,北京100853 [2]北京蛋白质组研究中心,蛋白质组学国家重点实验室,北京102206

出　　处：《现代检验医学杂志》2015年第4期14-20,共7页Journal of Modern Laboratory Medicine

基　　金：北京市科委首都临床特色应用研究项目（z121107005112009）,蛋白质组学国家重点实验室开放课题（SKLP-0200907）.

摘　　要：目的：建立一个良好的粪便蛋白质提取、分析和鉴定的方法，为进一步探索消化系统疾病无创性诊治靶标提供新思路和奠定良好研究体系。方法采用生理盐水、Tris-HCl 和尿素缓冲液三种粪便蛋白质提取体系，处理健康人群、慢性萎缩性胃炎、胃腺癌术前、胃腺癌术后的粪便标本，通过 SDS-PAGE 进行比较，获得较好的粪便蛋白质提取方法。再利用液相串联质谱（1D LC-MS／MS）对提取的蛋白质进行初步分析。结果生理盐水和 Tris-HCl 缓冲液提取粪便蛋白质的效率明显较尿素法高，损失少，是粪便蛋白质比较有效和合理的提取方法。不同人群来源的粪便中均含有丰富的蛋白质，正常人、胃腺癌手术前及术后鉴定到的蛋白质达一百多种，在组成和含量上个体差异明显。疾病与正常间、疾病治疗前后，粪便蛋白质指纹图谱变化大。胃腺癌术前粪便中含有大量的免疫球蛋白和角蛋白，同时也鉴定到了α1-抗胰蛋白酶，但在其术后和健康者粪便中未见到这一现象。结论正常与胃癌、胃癌手术前与术后粪便蛋白质不仅有蛋白质种类的差异，而且相同蛋白质量的差异也相当明显。因此，利用蛋白质组学技术筛选人粪便中胃癌相关的蛋白质标志物有较强的可行性，但在未来的研究中需要差异分析结合定量质谱的方法进行。Objective To set up a method of stools protein extraction,analysis and identification in order to get the new nonin-vasive indicators of human digestive diseases.Methods The stools proteins,collected from healthy persons,the patients with atrophic gastritis,those who suffed from gastric carcinoma and postoperative patients with gastric carcinoma respectively, were extracted in three different ways including saline,Tris-HCl buffer and Urea buffer,the best way was selected by using SDS-PAGE,then a preliminary analysis of stools proteins was performed by 1D LC-MS/MS.Results The methods of saline and Tris-HCl buffer could get more stools proteins than the method of urea.The proteins in stools from the healthy persons, the patients with atrophic gastritis,the patients with gastric carcinoma and postoperative patients with gastric carcinoma were all abundant and more than one hundred.There was a significant difference in stools protein maps among the various populations.Alpha1-antitrypsin,a number of immunoglobulin and keratin were identified in the stools from patients with gastric carcinoma but not postoperative patients with gastric carcinoma and the healthy persons.Conclusions In this re-search,there was a significant difference in stools protein maps among the healthy persons,the patients with atrophic gastri-tis,the patients with gastric carcinoma and postoperative patients with gastric carcinoma,not only the composition of stools proteins,but also the abundance of same proteins.Therefore,using proteomics technologies to screening of the noninvasive indicators in human stools is viable.The study recommended that the noninvasive indicators in human stools should be iden-tified with quantitative differences analysis combination of quality of mass spectrometer method in the future research.

关 键 词：粪便 蛋白质 胃腺癌 质谱 

分 类 号：R735.2[医药卫生—肿瘤] R730.43[医药卫生—临床医学]