应用二维电泳和质谱技术筛选大肠癌与正常肠组织的差异表达蛋白
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Screening of differential expression protein from human colorectal carcinoma and normal colorectal tissues by twodimensional difference gel electrophoresis and mass spectrometry
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    目的:通过筛选大肠癌组织与正常癌组织中的差异表达蛋白,以发现用于大肠癌早期诊断的生物标志物。方法:收集大肠癌组织及配对的正常肠组织,提取组织蛋白,进行二维电泳,选择在癌组织中高表达的30个点进行质谱和生物信息学分析,确定所分析的蛋白质类型。结果:正常肠组织和大肠癌组织的二维电泳图谱的平均蛋白质点数分别为760±45和1 098±28;正常肠组织在IEF方向上的平均偏差为(0.542±0.12)mm, 在SDSPAGE方向上的平均偏差为(0.933±0.098)mm;大肠癌组在IEF方向上的平均偏差为(0.745±0.130)mm,在SDSPAGE方向上的平均偏差为(1.233±0.272)mm。 对选择的30个在大肠癌中高表达的蛋白质点进行质谱和生物信息学分析,鉴定了其中的16个点,包括有Apolipoprotein A1,calreticulin precursor,谷胱苷肽S转移酶,肝型脂肪酸结合蛋白、热休克蛋白27等。结论:大肠癌与正常癌组织的差异表达蛋白可能作为大肠癌早期诊断的候选生物标志物;蛋白质组学技术是筛选生物标志物的有效方法。

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    ObjectiveTo find early diagnostic biomarkers for colorectal carcinoma by comparing differential expressingproteins from colorectal carcinoma and normal colorectal tissues. MethodsColorectal carcinoma tissues and paired normal tumoradjacent colorectal tissues were collected, and tissue total protein was extracted; differential proteome profiles were established and analysed by means of immobilized pH gradientbasedtwodimesional polyacrylamide gel electrophoresis(2DPAGE) and matrixassisted laser desorption/ionization time of flight mass spectrometry(MALDITOFMS). ResultsWellresolved, reproducible2DE profiles of human colorectal carcinoma tissues and paired normal tumor adjacent colorectal tissues were obtained. For tumor tissue, a total of 1098±28 spots were detected, and for normal tissue, 760±45 spots were detected. For normal tissue, The average deviation of spot position was(0.542±0.12)mm in IEF direction and(0.933±0.098)mm in SDSPGE direction for tumor tissue. The average deviation of spot position was( 0.745±0.130)mm in IEF direction and(1.233±0.272)mm in SDSPGEdirection. 30 differential expressing proteins were analysed by mass spectrometry and bioinformation,
    16 of them were well characterized including Apolipoprotein A1 (apoA1),calreticulin precursor,glutathione Stransferase, hepatic fatty acidbinding protein、heat shock protein 27 ect. ConclusionsDifferential expression proteins can be candidate biomarkers for early diagnosis of colorectal carcinoma; and proteomic technique is valuable for screening the diagnostic biomarkers.

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裴海平,朱红,曾亮,李宜雄.应用二维电泳和质谱技术筛选大肠癌与正常肠组织的差异表达蛋白[J].中国普通外科杂志,2005,14(10):10-752.
DOI:10.7659/j. issn.1005-6947.2005.10.010

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  • 在线发布日期: 2005-10-25