用激光捕获显微切割联合蛋白质芯片筛选肺鳞癌和腺癌组织差异蛋白  

Use of laser capture microclissection and surface-enhanced laser desorption ionization time-of-flight mass spectrometry to screen differential proteins in lung adenocarcinoma and lung squamous carcinoma

作　　者：田应选[1] 杨拴盈[1] 南岩东[1] 张潍[2] 周斌[2] 卜丽娜[1] 霍树芬[1] 余捷凯[3] 郑树[3] 

机构地区：[1]西安交通大学医学院第二附属医院呼吸科,710004 [2]西安交通大学医学院第二附属医院胸外科,710004 [3]浙江大学医学院附属第二医院肿瘤研究所

出　　处：《中华医学杂志》2008年第3期145-148,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目（30570795）;教育部新世纪优秀人才支持计划项目（NECT-06-0845）

摘　　要：目的应用激光捕获显微切割（LCM）联合表面增强激光解吸离子化飞行时间质谱（SELDI-TOF-MS）蛋白质芯片及支持向量机（SVM）方法筛选肺鳞癌和腺癌差异表达蛋白质，探讨二者在蛋白水平的差异，为筛选肺癌分型标志物提供依据。方法将6例新鲜肺鳞癌及7例腺癌组织标本用LCM选择性获取1．4×10^5个同质鳞癌细胞和1．2×10^5个同质腺癌细胞。经PBS Ⅱ^＋型SELDI—TOF-MS分析仪（IMAC 芯片）分析鳞癌及腺癌细胞蛋白质表达谱，比对差异峰；应用SVM筛选并验证候选标志蛋白的判别效能。结果比较鳞癌和腺癌细胞的SELDI谱图，共筛选出87个蛋白峰。将差异最明显的10个蛋白峰作为候选标志蛋白。与腺癌相比，4种蛋白（相对分子质量分别为2505、4004、4847及11 412）在鳞癌中呈高表达；与鳞癌相比，6种蛋白（相对分子质量分别为3333、3592、3848、5036、5191及5211）在腺癌中呈高表达。其中相对分子质量为4847的蛋白在鳞癌和腺癌中表达差异有统计学意义。用SVM建立分类预测模型并评价各模型效能，筛选出一个由3种蛋白质（相对分子质量分别为4847、11412和3592）组成的分型标志蛋白组合模式，其敏感度和特异度均为100％。结论肺鳞癌和腺癌在蛋白水平存在差异；LCM联合SELDI蛋白质芯片技术有可能筛选出敏感性高、特异性强的肺癌分型标志蛋白组合模式。Objective To screen biomarkers for classification in lung adenocarcinoma and lung squamous carcinoma by using laser capture microdissection （LCM） and surface-enhanced laser desorption ionization time-of-flight mass spectrometry （SELDI-TOF-MS） and support vector machine （SVM）. Methods Six specimens of lung adenocarcinoma tissues and seven specimens of lung squamous carcinoma tissues obtained during operation were made into frozen sections and stained by improved H-E solution. About 1.2 × 10^5 of homogeneous adenocarcinoma cells and 1.4 × 10^5 of homogeneous lung squamous carcinoma cells were collected using LCM. Then SELDI profiles based on PBS Ⅱ ^＋ SELDI-TOF-MS （IMAC protein chip） and the data were analyzed by support vector machine （SVM）. Results Eighty seven differential protein peaks were found and top ten of them were identified as candidate biomarkers. The expression levels of 6 proteins among them with the molecular weights of 3333, 3592, 3848,5036,5191, and 5211 respectively in the lung squamous cancer tissues were weaker than those in the adenocarcinoma tissues, and the expression levels of 4 proteins with the molecular weights of 2505,4004,4847, and 11 412 in the lung squamous carcinoma tissues were stronger than those in the adenocarcinoma tissues. The expression of the protein with the molecular weight of 4847 in the squamous cancer was significantly stronger than that in the adenocarcinoma （p ＋ 0. 032）. A discriminatory pattern consisting of 3 proteins with the molecular weights of 4847, 11 412, and 3592 was established with a sensitivity of 100% and a specificity of 100% respectively in separating adenocarcinoma from squamous carcinoma. Conclusion There is a difference in protein component between adenocarcinoma and squamous carcinoma. LCM combined with SELDI-TOF-MS help screen a biomarker pattern to distinguish lung adenocarcinoma from lung squamous carcinoma with high sensitivity and specificity.

关 键 词：光谱法 质量 基质辅助激光解吸电离 蛋白质组 肺肿瘤 激光捕获显微切割 

分 类 号：R686[医药卫生—骨科学]