比较蛋白质组学技术筛选葡萄膜黑色素瘤相关蛋白

Cancer-related proteins discovery of uveal melanoma with comparative proteomic analysis

目的应用比较蛋白质组学技术,分析鉴定葡萄膜黑色素瘤与正常人葡萄膜差异蛋白质,筛选出葡萄膜黑色素瘤相关蛋白。设计实验性研究。研究对象手术中切除的人葡萄膜黑色素瘤标本4例以及正常人捐献眼葡萄膜标本4例。方法研磨组织提取蛋白质后,应用双向凝胶电泳进行总蛋白分离,考马斯亮兰染色,Image Master 5.0软件分析蛋白胶图。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定部分差异蛋白质点,通过MASCOT软件在NCBI数据库中检索蛋白质信息。主要指标差异蛋白质。结果建立了葡萄膜黑色素瘤和正常人葡萄膜组织的双向凝胶电泳图谱,鉴定了葡萄膜黑色素瘤与正常人葡萄膜之间差异表达的蛋白质30种,其中24种蛋白只在葡萄膜黑色素瘤中出现。葡萄膜黑色素瘤中表达上调的蛋白5种,表达下调的蛋白1种。这些差异蛋白可能执行新陈代谢酶、信号转导调节、细胞骨架、免疫调节等生物学功能。结论通过比较蛋白质组学初步筛选出与葡萄膜黑色素瘤发生演进相关的蛋白质。需要进一步验证蛋白的定位及功能。

Objective To identify and characterize uveal melanoma associated protein variants with two-dimensional eleetrophoresis and mass spectrometry. Design Experiment study. Participants 4 cases of specimens of uveal melanoma and 4 cases of normal contributed uveal tissue. Methods Proteins from uveal melanoma and normal uvea were separated with two-dimensional electrophoresis （2-DE） and visualized with Coomassie G-250. Gels were analyzed by Image Master 5.0 software. The mass spectra were measured by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF MS） and searched against NCBI database using Mascot software. Main Outcome Measures Differential proteins. Results A set of 30 proteins were differentially expressed in uveal melanoma compared to nomal uvea. Twenty-four types of protein only expressed in uveal melanoma. Five types of protein were up-regulated and 1 type of one down-regulated. These proteins can be subdivided into groups according to cellular function, such as enzyme, signal transduction, signal regulation, cytoskeleton, immune, etc. Conclusions There is significant difference in protein profilings between uveal melanoma and normal uvea. The differentially expressed proteins may be associated with the development of uveal melanoma.