人胃癌原发灶与淋巴结转移灶的差异蛋白质组学研究  

Proteomics of differentially-expressed proteins between human primary and lymph node metastatic gastric adenocarcinoma

作　　者：柴丽丽[1] 陈燕[1] 王娟红[1] 郑长黎[2] 

机构地区：[1]西安市中心医院病理科,710003 [2]中南大学湘雅医学院病理学系

出　　处：《肿瘤研究与临床》2015年第9期586-588,592,共4页Cancer Research and Clinic

基　　金：国家自然科学基金（81372857）

摘　　要：目的：利用比较蛋白质组学技术识别并鉴定胃癌原发灶与淋巴结转移灶中的差异表达蛋白质，筛选特异性的转移相关蛋白，探讨胃癌淋巴结转移的分子机制。方法将前期应用二维差异凝胶电泳技术（DIGE）获取人胃癌原发灶及转移灶差异蛋白表达图谱进行分析获得的11个差异表达蛋白质，运用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）进行鉴定，得到相应的肽质量指纹图（PMF），然后搜索数据库鉴定出部分蛋白质。利用免疫组织化学技术检测HSP 70在胃癌中的表达水平。结果对11个差异蛋白质点进行PMF分析后，通过Mascot软件查询鉴定出其中5个蛋白质点（HSP70的8同种型2变异体、2个伴侣蛋白、亮氨酸氨肽酶、预测假想蛋白XP_515584）。 HSP70、伴侣蛋白、亮氨酸氨肽酶的表达在原发癌灶中明显升高。免疫组织化学检测示HSP 70的表达水平随着胃癌恶性程度的增加及淋巴结的转移逐渐增加，与蛋白质组学的分析结果一致。结论人胃癌原发灶与淋巴结转移灶中腺癌细胞的差异蛋白表达图谱相似，说明两者癌细胞的来源、分化程度相对一致，仅有少数蛋白质的质和量发生了明显变化，参与了淋巴结转移。其涉及细胞生理活动的各方面，与肿瘤的发生、发展等密切相关。 HSP70参与了胃癌的发生发展，其表达水平与胃癌有无淋巴结转移及恶性程度有密切关系。Objective To identify the differentially-expressed proteins of human gastric adenocarcinoma cell in primary or metastatic lymph node tissues by comparative proteomics technology, and to screen the specific metastatic-associated proteins so as to investigate the metastatic molecular mechanism of lymph node metastasis in gastric cancer. Methods 11differential proteins were acquired previously from primary and metastatic lymphnode tissues in gastric adenocarcinoma patients by 2D-DIGE. Some selected differential protein spots were identified by PMF based on MALDI-TOF-MS and database search. Immunohistochemical staining of HSP70 was used to evaluate the reliability of the proteomic analysis results. Results After analyzed on 11 differential proteins by in-gel trypsin digestion and MALDI-TOF-MS-based PMF analysis, a total of 5 differential proteins were identified by searching Mascot-database, HSP70ˊs 8 isoform 2 variant, chaperonin, chaperonin, leucine aminopeptidase, predicted： hypothetical protein XP_515584. Among the differential proteins identified, the levels of HSP70, chaperonin, leucine aminopeptidase expression had a significant up-regulation in gastric primary cancer compared with metastatic lymph node. HSP70 expression rate increased with the metastasis of lymph node and the progress of gastric cancer, agreed with the proteomics results. Conclusions They are similar in differentially-expressed proteins in primary or metastatic lymph node tissues because of the uniformity in source and differention. There are few protein changes in cancer cells between them, taking part in the metastatic of gastric cancer. HSP70 takes part in the progress of gastric cancer and relates to the metastasis of lymph node and malignant degree.

关 键 词：胃肿瘤 腺癌 淋巴转移 显微切割 光谱法 质量 基质辅助激光解吸电离 蛋白质组学 

分 类 号：R735.2[医药卫生—肿瘤]