蛋白质组学技术筛选肝癌血浆/血清中的差异蛋白质  

Selection and identification of differential proteins in plasma/serum with hepatocellular carcinoma by proteomic techniques

作　　者：尚京川[1] 易发平[2] 吴民军[3] 母昭德[1] 

机构地区：[1]重庆医科大学药学院药物分析教研室,重庆400016 [2]重庆医科大学分子医学与肿瘤研究中心,重庆400016 [3]重庆医科大学生命科学研究院,重庆400016

出　　处：《重庆医科大学学报》2010年第8期1212-1216,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金(编号:30800945);重庆市自然科学基金(编号:CSTC;2008BB5398)

摘　　要：目的:建立分析肝癌患者血浆/血清中低丰度差异表达蛋白质的双向凝胶电泳(2-DE)方法。方法:以10例肝癌患者血浆、8例肝癌患者血清及8例正常志愿者血浆和血清为研究对象,选用安捷伦免疫亲和色谱Agilent MARS Spin Cartridge去除血浆中6种主要高丰度蛋白,2-DE分离血浆/血清低丰度蛋白质,差异蛋白质斑点经酶解后,MALDI-TOF/MS检测的肽指纹谱(Peptide mass fingerprinting,PMF)在NCBInr数据库中进行搜索。结果:10例肝癌患者血浆样品中共鉴定出有20种差异蛋白,其中18种表达上调,2种表达。8例肝癌患者血清样品中共鉴定出23种差异蛋白,其中17种蛋白表达上调,6种蛋白表达下调。在肝癌患者血清和血浆组中共同识别到2种差异蛋白质。结论:血浆组中识别到的指环蛋白34(RING fingerprotein 34),威尔姆肿瘤1-关联蛋白(Wilms' tumor1-associating),CD147抗原,卵泡前体FS(Follistatin precursor,FS)蛋白和血清组中识别到的蛋白激酶NYD-SP9(Protein kinase NYD-SP9),肿瘤坏死因子受体超家族成员19前体(Tumor necrosis factor receptor superfamily member 19 precurso)r,组蛋白键合蛋白RBBP4(Histone-binding protein RBBP4),FADD蛋白,26S蛋白酶调控亚基P28(26S proteasome regulatory subunitp28),CD82抗原,自体吞噬蛋白5(Autophagyprotein5)等蛋白有望成为(Hepato celluar careinoma,HCC)的特异蛋白。Objective：To establish a two-dimensional polyacrylamide gel electrophoresi（s2-DE）technique forplasma/serumproteomics. Methods：Plasmas from 8 healthy volunteers and 10 with hepatocellular carcinoma（HCC）,and sera from 8 HCC were collected.After six proteins of the highest abundance in plasma/sera were depleted by the Multiple Affinity Removal System（MARS）Technologies from Agilent,plasma/sera were subjected to 2-DE.After comparing,analyzing and identifying by the software PD Quest（7.4,0）,the enzyme-digested products of differential expression proteins were analyzed by MALDI-TOF-MS.The Peptide mass fingerprinting（PMF） data were obtained and used to identify proteins through NCBInr data bank.Results：In comparison with the healthy volunteers,20 differential proteins were identified in the plasmas of the HCC patients,among which 18 protein expressions were up-regulated and 2 down-regulated.In the sera of the cancer patients,23 differential proteins were identified,among which 17 protein expressions were up-regulated and 6 down-regulated.Tumor necrosis factor-inducible protein TSG-14 and FADD protein were both identified in the plasmas and sera of the cancer patients.Conclusion：The differential proteins,including RING finger protein 34,Wilms’tumor 1-associating protein,CD147 antigen,Tumor necrosis factor-inducible protein TSG-14 from plasmas group and Protein kinase NYD-SP9,Tumor necrosis factor receptor super family member 19 precursor,Histone-binding protein RBBP4,FADD protein,26S protease regulatory subunit p28,CD82 antigen,Autophagy protein 5 from sera group may be used as markers for HCC in blood.

关 键 词：肝细胞癌 血浆/血清 蛋白质组学 双向电泳 MALDI-TOF/MS 

分 类 号：Q592.1[生物学—生物化学] R730.2[医药卫生—肿瘤]