胶质瘤组织中蛋白质表达谱的研究及疾病相关蛋白质的筛选

Research on Glioma Protein Profile and Screening of Its Disease-related Proteins in Tissues

目的利用双向凝胶电泳-MALDI-TOF质谱技术进行胶质瘤组织中蛋白质表达谱的研究,并进行胶质瘤组织中疾病相关蛋白质的筛选。方法分别提取6例胶质瘤患者脑组织、6例相对正常脑组织的总蛋白质,进行双向凝胶电泳、银染,两组凝胶图像差异分析,得到差异蛋白质。对目的差异蛋白质进行MALDI-TOF质谱分析。结果获得了分辨率较高的胶质瘤和相对正常脑组织双向凝胶电泳图谱,鉴定了29个差异蛋白质点中的18个蛋白质,得到15个阳性结果。在鉴定中得到阳性结果的差异蛋白质中,胶质瘤组织上调的蛋白质有:γ-肌动蛋白、三联基元蛋白5的环指蛋白、假想蛋白、RAS结合蛋白、T细胞识别的鳞状细胞癌抗原、SMC1结构维持染色体1样1、ATP结合盒转运载体蛋白1和果糖二磷酸醛缩酶;下调的蛋白质有:β-球蛋白、脑肌酸激酶、类似S期激酶活化剂、CUG-BP和ETR-3样因子、类似磷酸二酯酶4D相互作用蛋白、CD2抗原和肌动蛋白结合蛋白。结论利用双向凝胶电泳-MALDI-TOF质谱技术进行胶质瘤组织中疾病相关蛋白质的研究得到差异蛋白质,其中上调蛋白质功能为细胞的骨架结构形成、细胞的侵袭性生长、肿瘤耐药性和促进细胞增殖等;下调蛋白质功能为免疫调节、细胞周期调节、抑制细胞迁移、在脑组织中参加细胞能量代谢和调节细胞特异性和发育等;还有一些未知功能蛋白质。所得结果为胶质瘤疾病蛋白质组学的进一步研究打下了一定的工作基础。

Objective To study glioma protein profile and to screen its disease - related proteins in tissues by two - dimensional gel electrophoresis （2DE） - MALDI - TOF mass spectrometry. Methods Total proteins were extracted from brain tissues of 6 glioma patients and 6 normal subjects, and then 2DE and silver staining were done, the differences in gel image between the two were analyzed, and differential proteins were then obtained, on which MALDI - TOF mass spectrometry analysis was performed. Results We got a high - resolution 2DE of brain tissues, identified 18 of 29 differential proteins and got positive results from 15. In positive differential proteins, the up -regulated proteins included gamma -actin, ring finger protein of tripartite motif protein （ TRIM5 ）, hypothetical protein, RAS - binding protein, squamous cell carcinoma antigen recognized by T -cell, SMC1 structure maintenance chromosome 1 -like 1, ATP -binding cassette transportation carrier protein - 1 and fructose - bisphosphate aldolase C ; the down - regulated proteins included beta globulin, brain creatine kinase, activator similar to S -phase kinase, CUG- BP and ETR -3 -like factor, 4D interactive protein similar to phosphodiesterase, CD2 antigen and actin - binding protein. Some of them were hypothetical proteins. Conclusion Use of 2DE - MALDI - TOF mass spectrometry in research of disease - related proteins in glioma tissues can help get differential proteins, of which up - regulated proteins have functions of formation of cell backbone structure, invasive cell growth, drug resistance of tumor, promotion of cell proliferation, and down - regulated proteins function as immune regulation, cell cycle regulation, inhibition of cell migration, participation in cell energy metabolism and regulation of cell specificity and development in brain tissue. Of course there are still some proteins whose functions are unknown to us. The results which have been got will lay basis for further research of glioma disease proteomics.