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凝溶胶蛋白作为中国人群结直肠癌潜在生物标志物的发现与验证:采用基于iTRAQ标记的蛋白质组学方法检测差异蛋白表达

Discovery and verification of gelsolin as a potential biomarker of colorectal adenocarcinoma in the Chinese population: Examining differential protein expression using an iTRAQ labelling-based proteomics approach.

作者信息

Fan Nai-Jun, Gao Chun-Fang, Wang Chang-Song, Lv Jing-Jing, Zhao Guang, Sheng Xin-Hua, Wang Xiu-Li, Li Dong-Hui, Liu Qing-Yin, Yin Jian

机构信息

Anal-Colorectal Surgery Institute, LuoYang, China.

出版信息

Can J Gastroenterol. 2012 Jan;26(1):41-7. doi: 10.1155/2012/645218.

Abstract

OBJECTIVE

To identify and validate potential biomarkers of colorectal adenocarcinoma using a proteomic approach.

METHODS

Multidimensional liquid chromatography⁄mass spectrometry was used to analyze biological samples labelled with isobaric mass tags for relative and absolute quantitation to identify differentially expressed proteins in human colorectal adenocarcinoma and paired normal mucosa for the discovery of cancerous biomarkers. Cancerous and noncancerous samples were compared using online and offline separation. Protein identification was performed using mass spectrometry. The downregulation of gelsolin protein in colorectal adenocarcinoma samples was confirmed by Western blot analysis and validated using immunohistochemistry.

RESULTS

A total of 802 nonredundant proteins were identified in colorectal adenocarcinoma samples, 82 of which fell outside the expression range of 0.8 to 1.2, and were considered to be potential cancer-specific proteins. Immunohistochemistry revealed a complete absence of gelsolin expression in 86.89% of samples and a reduction of expression in 13.11% of samples, yielding a sensitivity of 86.89% and a specificity of 100% for distinguishing colorectal adenocarcinoma from normal tissue.

CONCLUSIONS

These findings suggest that decreased expression of gelsolin is a potential biomarker of colorectal adenocarcinoma.

摘要

目的

采用蛋白质组学方法鉴定并验证结直肠癌的潜在生物标志物。

方法

运用多维液相色谱/质谱联用技术,对用等压质量标签标记的生物样品进行相对和绝对定量分析,以鉴定人类结直肠癌及配对的正常黏膜中差异表达的蛋白质,从而发现癌生物标志物。使用在线和离线分离方法比较癌组织和非癌组织样品。通过质谱进行蛋白质鉴定。采用蛋白质印迹分析确认结直肠癌样品中凝溶胶蛋白的下调,并通过免疫组织化学进行验证。

结果

在结直肠癌样品中总共鉴定出802种非冗余蛋白质,其中82种蛋白质的表达范围不在0.8至1.2之间,被认为是潜在的癌症特异性蛋白质。免疫组织化学显示,86.89%的样品中完全没有凝溶胶蛋白表达,13.11%的样品中表达减少,区分结直肠癌与正常组织的灵敏度为86.89%,特异性为100%。

结论

这些发现表明,凝溶胶蛋白表达降低是结直肠癌的一种潜在生物标志物。

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