鼻息肉的2-DE图谱建立和蛋白质组学分析  

Establishment of 2-dimensional gel electrophoresis map and analysis of proteomics from human nasal polyps

作　　者：贺广湘[1] 孙虹[1] 王天生[1] 李萃[2] 刘火旺[1] 陈玉[1] 

机构地区：[1]中南大学湘雅三医院耳鼻咽喉科,长沙410013 [2]中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室,长沙410008

出　　处：《中南大学学报（医学版）》2006年第4期487-492,共6页Journal of Central South University ：Medical Science

基　　金：教育部博士点基金(20030533006);湖南省卫生厅课题(B2004-069);中南大学博士创新选题(030639)

摘　　要：目的:建立鼻息肉及鼻黏膜双向凝胶电泳(two-dimensional polyacrylamide gel electrophoresis,2-DE)图谱,鉴定差异表达蛋白质。方法:收集鼻息肉及鼻黏膜样本各7例,采用固相pH梯度2-DE,凝胶银染,扫描图像,ImageMaster2-DE软件比较分析等方法,识别差异表达蛋白质,通过质谱分析得到相应肽质指纹图谱(peptide mass fingerprint,PMF),采用PeptIdent软件查询Swiss-Protand TreMBL数据库,鉴定差异表达蛋白质。结果:建立了鼻息肉和鼻黏膜蛋白质的2-DE图谱。鼻息肉和鼻黏膜3块凝胶的平均蛋白质点数分别为825±78和936±62;平均匹配点数为682±96和821±78,匹配百分率为82.7%和87.7%;同一鼻息肉的3块胶在蛋白质点位置上有较好的重复性,不同胶间蛋白质点在等电聚焦(IEF)方向偏差为(1.06±0.14)mm,在SDS-PAGE方向偏差为(1.45±0.21)mm。比较分析两种组织各7例样本的2-DE图谱,鼻息肉和鼻黏膜蛋白质点数为1458个和1617个,平均匹配点数为1026个。质谱分析差异蛋白质点40个,获取34张PMF,查询数据库鉴定出蛋白质24个。结论:建立了分辨率高和重复性好的鼻息肉及鼻黏膜2-DE图谱,识别鉴定出一些与鼻息肉病变相关的蛋白质。Objective To establish 2-dimensional polyacrylamide gel electrophoresis （2-DE） map from human nasal polyps and normal nasal mucosa, and to identify differential expression proteins of 2-DE map. Methods Samples of nasal polyps and nasal mucosa （ each sample group containing 7 cases ） were obtained. The total proteins were extracted and separated by immobilized pH gradient （IPG）-based 2-DE. The silver-stained 2-DE was scanned with digital lmagescanner and analyzed with ImageMaster 2-DE Elite 4.01 software. To obtain peptide mass fingerprint （ PMF ） of differential protein spots, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS） was used. The PMF was searched in Swiss-Prot and TreMBL database by Peptldent software, to identify differential expression proteins. Results The well-resolved, reproducible 2-DE maps of nasal polyps and nasal mucosa were established. For the polyps tissues, the average proteins spot of three 2-DE maps was 825 ± 78 ; and 682 ± 96 spot was matched with the average matching rate of 82.7 % . The average deviations of matched spot position were （ 1. 13±0. 16 ） mm in IEF direction and （ 1.45±0.21 ） mm in SDS-PAGE direction, respectively. For the nasal mucosa tissues, the average proteins spot of three 2-DE maps was 936 ± 62; and 821±78 spots were matched with the average matching rate of 87.7%. After comparing the 2-DE maps of nasal polyps and nasal mucosa tissues, the protein spots were 1 458 and 1 617 respectively; and 1 026 protein spots were matched. Forty differential expression protein spots were incised from sliver staining gel randomly and digested in the gel by TPCK-Trypsin. Thirty-four PMFs were obtained by MALDI- TOF-MS and 24 differential proteins were identified. Conclusion The well-resolved, reproducible 2-DE maps of human nasal polyps and nasal mucosa have been successfully established. Certain dif- ferential proteins related to the pathogenesis of human nasal polyps are identified.

关 键 词：鼻息肉 鼻黏膜 蛋白质组学 差异表达蛋白质 

分 类 号：R765.25[医药卫生—耳鼻咽喉科]