利用激光捕获和高效液相色谱质谱对石蜡包埋口腔黏膜癌变组织的蛋白组学分析  

A proteomics study of oral mucosa carcinogenesis basing on laser capture microdissection and liquid chromatographymass spectrometry

作　　者：耿宁[1,2] 王睿男[1,3] 周玉乔[1,4] 张敦房[1] 张乐薇 吉宁[1] 周敏[1] 梁新华[1] 陈宇[1,2] 李敬[1] 陈谦明[1,4] 

机构地区：[1]口腔疾病研究国家重点实验室,四川成都610041 [2]四川大学华西口腔医院病理科,四川成都610041 [3]农业部中国动物疫病预防控制中心,北京100125 [4]四川大学华西口腔医院黏膜病科,四川成都610041 [5]加拿大英属哥伦比亚大学牙学院肿瘤研究中心

出　　处：《口腔生物医学》2015年第4期188-192,共5页Oral Biomedicine

基　　金：国家自然科学基金国际合作交流中加项目(81010066)

摘　　要：目的：对口腔黏膜癌变的蛋白组学分析和分子标志物筛选可能为早期诊断、预防和分子治疗提供依据和帮助。方法：对3例石蜡包埋口腔黏膜癌变组织样本,利用激光显微切割技术捕获口腔黏膜癌变上皮和癌旁正常黏膜上皮,抽提组织样本蛋白,利用高效液相色谱-质谱联用进行口腔黏膜癌变组织的蛋白组学分析。结果：激光显微切割-高效液相色谱质谱联用可以分析400~700个多肽序列,鉴定出100~200个蛋白,涉及到细胞骨架、细胞代谢、信号转导、细胞周期等各个方面。结合文献,对其中的角蛋白表达变化进行了分析和验证。结论：显微切割结合高效液相色谱质谱可以实现对石蜡包埋组织的蛋白组学研究,具有较高的重复性和可靠性。角蛋白表达谱的变化验证了该平台的作用和价值,也可作为口腔黏膜癌变的潜在标志物。Objective： OSCC is associated with poor prognosis,which has improved only marginally over the past three decades. A proteomic analysis of OSCC lesions may help identify novel molecular targets for the early detection,prevention,and treatment of it.Methods： Laser capture microdissection was combined with recently developed techniques for protein extraction from formalin-fixed paraffin-embedded（ FFPE） tissues and a novel proteomics platform. Approximately 10,000 cells procured from FFPE tissue sections of normal oral epithelium and OSCC epithelium were processed for mass spectrometry and bioinformatic analysis. Results： A large number of proteins expressed in normal oral epithelium and OSCC,including cytokeratins,intermediate filaments,differentiation markers,and proteins involved in stem cell maintenance,signal transduction,migration,cell cycle regulation,growth and angiogenesis,matrix degradation,and proteins with tumor suppressive and oncogenic potential,were readily detected. Of interest,the relative expression of many of these molecules followed a distinct pattern in normal squamous epithelia and OSCC tumor tissues. Representative proteins were further validated basing on article reviews. CKs expression pattern may afford potential predict biomarkers for oral mucosa carcinogenesis. Conclusions： The ability to combine laser capture microdissection and in-depth proteomic analysis of FFPE tissues provided a wealth of information regarding the nature of the proteins expressed in normal squamous epithelium and during OSCC progression,which may allow the development of novel biomarkers of diagnostic and prognostic value and the identification of novel targets for therapeutic intervention in OSCC.

关 键 词：口腔癌 石蜡包埋组织 蛋白组学 质谱 激光显微切割 

分 类 号：R780.2[医药卫生—口腔医学]