【作者】 胡柳； 张喜梅； 周福祥； 周云峰；

【Author】 HU liu,ZHANG Xi-mei,ZHOU Fu-xiang,ZHOU Yun-feng Department of Radio-chemotherapy,Zhongnan Hospital,Wuhan University,Whan 430071

【机构】 武汉大学中南医院肿瘤放化疗科；

【摘要】 目的在成功建立的辐射诱导放射抗拒鳞癌细胞株的基础上,以蛋白质组学分析方法筛选出放射抗拒与放射敏感鳞癌细胞株的差异蛋白蛋白质,探讨放射敏感性的调控机制,寻找新的放射增敏靶点。方法以双向电泳建立同一遗传背景的人喉鳞癌细胞株(Hep-2)及放射抗拒细胞株(Hep-2R)的全蛋白银染电泳图谱,以imageMasterTM 2D version:5.0图像分析软件对两细胞株电泳图比对分析,综合人工比对及软件比对结果,挑选出差异表达≥1.5倍(P<0.01)的蛋白斑点,另以考染胶作为制备胶,将差异蛋白从考染胶上切点、胶内酶解后,以Synapt HDMS离子迁移质谱仪分析并搜索NCBInr数据库。结果通过Hep-2及Hep-2R细胞株双向电泳图谱,筛选并鉴定出磷酸甘油酸酯激酶1、酸性磷酸酶同工酶、泛素结合酶、8型17-beta羟基固醇去氢酶、磷酸丙糖异构酶、黄曲霉毒素乙醛还原酶、ADP核糖基化因子相互作用蛋白7个在Hep-2R细胞株表达下调蛋白,以及丝切蛋白、过氧化物酶3、监控蛋白10、醌还原酶、线粒体DNA结合蛋白、磷酸核糖焦磷酸合成酶、人高迁移率族蛋白1(HMG-1)7个上调的蛋白。结论这些差异蛋白质与放射敏感性存在密切的关系。它们分别参与了物质代谢、DNA损伤修复、细胞周期调控等过程,说明放射敏感性是复杂的、由多途径调控的。更多还原

【Abstract】 Objective To find the different proteins in radioresistant and radiosensitive squamous carcinoma cell by comparative proteomic analysis,to investigate the mechanisms of radioresistance.Methods Our colleague had established and isolated the radioresistant cell line Hep-2R from the radiosensitive human larynx squamous carcinoma cell line Hep-2.The whole proteins from Hep-2 and Hep-2R were separated by two-dimensional gel electrophoresis.Silver stained gels were used for comparative analysis by using ImageMaster 2D platinum software. Proteins spots which differentially expressed more than 1.5 fold were chose for spectrometer analysis.These proteins were picked up in Coomassie brilliant blue-stained gel for spectrometer analysis by using Synapt HDMS mass spectrometer. All MS/MS data were probed in the NCBInr database by using the Mascot search engine. Results The fourteen proteins which differentially expressed more than 1.5 fold were found in gels(P<0.01).Seven proteins which up-express in Hep-2R cell line were identified as peroxiredoxin 3,chaperonin 10,Human Quinone Reductase Type 2, Human Mitochondrial Single-Stranded Dna Binding Protein,phosphoribosyl pyrophosphate synthetase 1 and HMG-1.There are seven down-expressing proteins in Hep-2R,including acid phosphatase isoenzyme,cofilin 1,ubiquitin-conjugating enzyme E2N,Triosephosphate Isomerasel,estradiol 17 beta-dehydrogenase 8,phosphoglycerate kinase 1 and aflatoxin.Conclusion These proteins may have a close relation with radioresistance in Hela-R cell by involving the process of metabolism,DNA damage repair and cell cycle,which suggests that the regulation of radioresisitance is a complex and multi-step process.更多还原