摘要
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3 0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7 6 0±4 5和1 0 9 8±2 8;正常肠组织在IEF方向上的平均偏差为(0.5 4 2±0.1 2)mm,在SDS-PAGE方向上的平均偏差为(0.9 3 3±0.098)mm;大肠癌组在IEF方向上的平均偏差为(0.7 4 5±0.1 3 0)mm,在SDS-PAGE方向上的平均偏差为(1.2 3 3±0.2 7 2)mm。对选择的3 0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1 6个点,包括有Apolipoprotein A 1,calreticulin precursor,谷胱苷肽S转移酶,肝型脂肪酸结合蛋白、热休克蛋白2 7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。
Objective To find early diagnostic biomarkers for colorectal carcinoma by comparing differential expressing proteins from colorectal carcinoma and normal colorectal tissues. Methods Colorectal 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-based two-dimesional polyacrylamide gel electrophoresis （2D-PAGE） and matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS）. Results Well-resolved, reproducible 2-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-PGE direction. 30 differential expressing proteins were analysed by mass spectrometry and bioinformation,16 of them were well characterized including Apolipoprotein A 1 （ apoA 1 ） , calreticulin precursor, glutathione S-transferase, hepatic fatty acid-binding protein, heat shock protein 27 ect. Conclusions Differential expression proteins can be candidate biomarkers for early diagnosis of colorectal carcinoma ; and proteomic technique is valuable for screening the diagnostic biomarkers.
出处
《中国普通外科杂志》
CAS
CSCD
2005年第10期748-752,共5页
China Journal of General Surgery
基金
湖南省科技厅资助项目(04SK3043-1)
湖南省卫生厅资助项目(B2004-029B2004-129)
