放疗敏感性不同宫颈癌组织的蛋白质组学比较研究  

Comparative study on the proteomic analysis in cervical carcinoma tissues with different radiosensitivities

作　　者：曾亮[1] 朱红[2] 裴海平[3] 邓亚平[1] 袁君[2] 

机构地区：[1]湖南省肿瘤医院病理科,长沙410013 [2]中南大学湘雅医院肿瘤科,长沙410008 [3]中南大学湘雅医院普外科,长沙410008

出　　处：《肿瘤》2007年第10期808-812,共5页Tumor

基　　金：湖南省自然科学基金资助项目(编号:06JJ4199);湖南省卫生厅课题项目(编号:B2004-029)

摘　　要：目的:比较放疗敏感性不同的早期宫颈癌组织之间蛋白质组的差异,为确定宫颈癌放疗敏感性相关标志物提供依据。方法:收集进行治疗前的早期宫颈癌组织标本。根据放疗后WHO实体瘤疗效判断标准,选择其中对放疗敏感和不敏感的标本分为高敏感组和低敏感组。提取组织总蛋白,进行双向凝胶电泳得到凝胶图谱,采用PD-quest 7.0软件进行匹配和差异分析,识别两组之间表达差异蛋白点。将部分差异蛋白点进行胶内原位切割、酶解后进行MALDI-TOF-MS分析,获取肽质量指纹图,数据库搜索鉴定蛋白质。应用Western印迹技术验证筛选的部分差异表达蛋白的表达。结果:建立了分辨率高、重复性好的宫颈癌放疗高敏感组和低敏感组的双向凝胶电泳图谱。高敏感组蛋白点(754±64)个(n=5),低敏感组蛋白点(777±48)个(n=5),组间平均匹配率为87.6%。选择部分差异在2倍以上的蛋白斑点进行质谱分析,8个差异表达蛋白被成功鉴定,其中5个蛋白质在高敏感组高表达,3个蛋白质在高敏感组低表达。Western印迹结果与蛋白质组筛选结果基本一致。结论:放疗高敏感组和低敏感组宫颈癌组织间存在蛋白表达的差异,这些差异表达的蛋白可能与放疗敏感性有关。Objective： To compare the proteomic differences in early cervical carcinoma tissues with various radiosensitivities and provide the basis for determining of radiosensitivity-associated proteins in cervical carcinoma. Methods： The fresh carcinoma tissues were collected from 10 untreated cervical carcinoma patients and preserved at - 80℃. According to WHO Criterion of Therapeutical Effect of Solid Carcinoma, the tissues were classified into two groups： high sensitive group （HS） and low sensitive group （LS）. The total protein was extracted and separated by using two-dimensional gel electrophoresis （2-DE）. The PD-quest 7.0 software was used to perform image matching and difference analysis to identify the differentially expressed protein-spots between the two groups. The identified protein spot was cut in electrophoretic gel in situ, digested with trypsin, and analyzed by Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry （MALDI-TOF-MS）. The peptide mass fingerprintings （PMF） was obtained. The proteins were identified by data searching in the Mascot database. Some differentially expressed proteins were confirmed by Western blotting. Resuits： The constructed two-dimensional gel electrophoresis profiles had high resolution and good reproducibility for both HS and LS groups. There were （754±64） differentially expressed protein spots （n =5） in HS group and （777±48） in LS group （n =5）. Some protein spots with differential ratio more than 2-fold were selected and analyzed by mass spectrometry. The average matching rate was 87.6% between the two groups. Eight differentially expressed proteins were successfully identified among which five proteins had overexpression and three proteins had weak expression in HS group. The results of Western blotting were consistant with proteomic screening. Conclusion： There was significant difference in protein profilings between HS and LS early cervical carcinoma after radiotherapy. The differentially expressed prot

关 键 词：宫颈肿瘤 放射治疗 蛋白质组学 电泳 凝胶 双向 光谱分析 质量 

分 类 号：R737.33[医药卫生—肿瘤]