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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 compare differentially expressed proteins between uveal melanoma and normal human uvea using comparative proteomics and screen out the uveal melanoma related proteins. Design experimental research. Four cases of uveal melanoma resected in the study subjects and four cases of uveal uterine donation in normal subjects were included in the study. Methods After grinding the tissue to extract the protein, the total protein was separated by two-dimensional gel electrophoresis. Coomassie brilliant blue staining and Image Master 5.0 software were used to analyze the protein gum map. Partially differentially expressed protein spots were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), and protein information was retrieved in the NCBI database by the MASCOT software. The main indicators of differential proteins. Results Two-dimensional gel electrophoresis patterns of uveal uveal tissue and normal uveal tissue were established and 30 proteins were differentially expressed between uveal melanoma and normal uveal uvea. Among them, 24 proteins expressed only in uveal melanoma Appear. There are 5 kinds of proteins that are upregulated in uveal melanoma and 1 kinds of proteins are down-regulated. These differential proteins may perform biological functions such as metabolic enzymes, signal transduction regulation, cytoskeleton, immune regulation and the like. Conclusion Proteins related to the evolution of uveal melanoma were initially screened by comparing proteomics. Need to further verify the protein localization and function.