蛋白质组学方法鉴定喉鳞状细胞癌中的肿瘤相关蛋白

目的利用蛋白质组学方法，建立人喉鳞状细胞癌（简称鳞癌）组织及其癌旁正常黏膜组织的双向凝胶电泳图谱，并识别鉴定其差异表达的蛋白质。方法利用固相pH梯度分离人喉鳞癌组织及癌旁正常黏膜组织的总蛋白质，用图像分析软件比较，以识别差异表达的蛋白质；应用质谱仪得到相应的肽质指纹图谱，’然后搜索数据库鉴定部分差异蛋白质点。结果获得分辨率和重复性均较好的双向凝胶电泳图谱，并鉴定出可能与喉癌发生密切相关的13种蛋白质。其中有10种蛋白质在喉癌组织中上调，分别是切丝蛋白1（cofilin-1），核体蛋白SP140（nuclear body protein SP140），内质网素，热休克蛋白90（heatshockprotein90，HSP90），谷胱甘肽S转移酶P（glutathioneS-transferase，GSTP1—1），锰-超氧化歧化酶（superoxidedismutaseMn]），亲环素A（cyclophilinA），蛋白酶活性复合体2（proteasome activator complex subunit2），载脂蛋白A-I前体（apolipoproteinA．Iprecursor），钙调素（CaM—like protein）。3种蛋白质在喉癌组织中下调，分别是银屑病相关脂肪酸结合蛋白（fatty acidbinding protein，epidermal，E—FABP），钙粒蛋白A（calgranulin A），钙粒蛋白B（calgranulin B）。结论本研究鉴定出了可能与喉癌的发生密切相关的13种蛋白质，提示多种蛋白分子可能同时成为应对这种疾病的有效策略的靶分子，为理解喉癌的癌变机制和改善临床诊治方法提供理论基础。

Identification of tumor-associated proteins in laryngeal squamous cell carcinoma by proteomics

OBJECTIVE: To establish two-dimensional electrophoresis profiles from human laryngeal squamous cell carcinoma tissue and paired normal tumor-adjacent mucosa epithelia tissue, and to identify differential expression proteins. METHODS: The total proteins of human laryngeal squamous carcinoma tissue and paired normal tumor-adjacent mucosa epithelia tissue were separated by immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE). The differential expression proteins were analyzed using image analysis software, then identified using mass spectrometry and database searching. RESULTS: Well-resolved, reproducible 2-DE patterns of laryngeal squamous cell carcinoma and adjacent normal mucosa epithelial were obtained. Differential protein spots were defined as spots in 2-DE gels. Thirteen proteins were preliminarily identified, namong which 10 proteins were upregulated in laryngeal cancer tissue. Such as cofilin-1, nuclear body protein SP140, GRP94, HSP 90, GSTP1-1, superoxide dismutase Mn], cyclophilin A, proteasome activator complex subunit 2, apolipoprotein A-I precursor, CaM-like protein and so on. There were 3 proteins downregulated in laryngeal cancer tissue, which were fatty acid-binding protein, calgranulin A and calgranulin B. CONCLUSIONS: Thirteen proteins which are associated with the tumorigenesis of the laryngeal squamous cell carcinoma were characterized. These extensive protein variations indicate that multiple protein molecules should be simultaneously targeted as an effective strategy to counter the disease. It is better for understanding of the oncogenesis and pathogenesis in a global way, which in turn is a basis-for the rational designs of diagnostic and therapeutic methods.