摘要
目的:分析结直肠癌(colorectal cancer,CRC)与正常黏膜间的蛋白质表达差异,筛选新的肿瘤标志物;并对兴趣蛋白进行验证,分析其与CRC的发生、发展及淋巴结转移的关系.方法:对6对新鲜的CRC与正常黏膜组织进行以二维差异凝胶电泳(2D differential gel electrophoresis,2D DIGE)及基质辅助激光解吸飞行时间质谱(matrix-assisted laserde sorption/ionization-time of flight masss pectrometry,MALDI-TOF-MS)分析.以免疫组织化学法验证兴趣蛋白泛醌细胞色素c还原酶核心蛋白1(ubiquinol cytochrome-creductase core protein 1,UQCRC1)在78例CRC与正常黏膜组织,和24个转移淋巴结中的表达.对染色的强弱评分为阴性:0,弱阳性:1,中阳性:2,强阳性:3.结果:2DDIGE分析显示在CRC中一个蛋白点丰度平均显著增高2.14倍(P<0.001).MALDI-TOF-MS分析证实该蛋白为UQCRC1.免疫组织化学法分析显示UQCRC1在CRC与正常黏膜组织中表达分别为2.28±0.95和1.81±0.88,有显著差异(P<0.001).UQCRC1表达的强弱与分化、分期及部位均无关(P>0.05).UQCRC1在转移淋巴结与配对的原发灶中表达分别为2.79±0.51和2.33±0.96,有显著差异(P<0.05).结论:UQCRC1在结直肠癌变和淋巴结转移过程中发挥一定的作用.
AIM：To screen new biomarkers for colorectal cancer （CRC） by identifying differentially expressed proteins between CRC and matched normal mucosa using proteomic technology and to analyze the association of identified biomarkers with colorectal carcinogenesis,development and lymph node metastasis.METHODS：Six pairs of fresh samples of CRC and matched normal mucosa were analyzed by two-dimensional differential gel electrophoresis （2D-DIGE） followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS）.The expression of the identified differential protein ubiquinolcytochrome c reductase core protein 1 （UQCRC1） was validated in 78 cases of CRC,matched normal mucosa and lymph node metastases.The immunostaining intensity of UQCRC1 was scored.Wilcoxon signed rank test and KrukalWallis test were used for statistical analysis.RESULTS：2D-DIGE analysis showed that the intensity of a protein spot was significantly increased by 2.14 folds （P 0.001） in CRC.This spot was confirmed to be UQCRC1 by MALDITOF-MS.Immunohistochemistry analysis revealed that the immunostaining intensity of UQCRC1 was significantly higher in CRC than in matched normal mucosa （2.28 ± 0.95 vs 1.81 ± 0.88,P 0.001）.However,the expression of UQCRC1 had no correlation with differentiation,TNM stage or tumor location （all P 0.05）.The intensity of UQCRC1 was significantly lower in primary CRC than in lymph node metastases （P 0.05）.CONCLUSION：UQCRC1 may play an important role in colorectal carcinogenesis and lymph node metastases.
出处
《世界华人消化杂志》
CAS
北大核心
2012年第3期259-263,共5页
World Chinese Journal of Digestology
基金
辽宁省教育厅一般基金资助项目
No.L2010600~~
