鼻息肉病与正常鼻黏膜组织的蛋白质组差异分析

目的:建立分辨率高和重复性好的鼻息肉病及对照鼻黏膜双向凝胶电泳(2DE)图谱,筛选鉴定差异表达蛋白质。方法:收集鼻息肉病(鼻息肉病组)及对照鼻黏膜标本(对照组)各7例,固相pH梯度2DE、凝胶银染,扫描图像,ImageMaster2DEElite4.01软件分析,识别差异表达蛋白质,质谱仪得到相应肽质指纹图谱(PMF),PeptIdent软件查询SwissProtandTreMBL数据库,鉴定差异表达蛋白质。结果:①鼻息肉病组和对照组各自随机挑选的1个样本3次重复2DE,平均蛋白质点数分别为891±67和936±62;匹配点数为767±83和821±78,平均匹配百分率为86.1%和87.7%;同一鼻息肉病的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电泳(IEF)方向偏差为(1.13±0.16)mm,在SDS聚丙烯酰胺凝胶电泳(SDSPAGE)方向偏差为(1.45±0.21)mm。分析2种组织各7例样本的2DE图谱的平均凝胶图谱,鼻息肉病组和对照组蛋白质点数为1532和1617,匹配点数为1065个。②质谱分析差异蛋白质点20个,获取16张PMF,查询数据库鉴定出差异蛋白质11个。结论:本研究建立了分辨率高和重复性好的鼻息肉病及鼻黏膜的2DE图谱,识别鉴定出一些与鼻息肉病变相关的差异表达蛋白质。

Differential proteins analysis for human nasal polyposis and normal nasal mucosa

OBJECTIVE: To establish two-dimensional polyacrylamide gel electrophoresis (2-DE) map with high resolution and reproducibility from human nasal polyposis and normal nasal mucosa, and to identify differential expression proteins of 2-DE map. METHOD: Samples of human nasal polyposis and normal nasal mucosa (each sample group containing seven cases) were obtained. The total proteins were extracted and separated by immobilized pH gradient (IPG)-based 2-DE. The silver-stained 2-DE were scanned with digital Image Scanner and analyzed with ImageMaster 2-DE Elite 4.01 software. Peptide mass fingerprint (PMF) of differential protein spots was obtained with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The peptide mass fingerprints were searched in Swiss-Prot and TreMBL database by PeptIdent software, and then differential expression proteins were identified. RESULT: (1) 2-DE for a randomly selected 1 sample from each of the 2 groups was repeated 3 times respectively to analyze the reproductive of the method. The image analysis showed: For the polyposis tissues, the average proteins spots of three 2-DE maps were 891 +/- 67 and 767 +/- 83, spots were matched with the average matching rate of 86.1%. 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 spots of three 2-DE maps were 936 +/- 62 and 821 +/- 78, spots were matched with the average matching rate of 87.7%. Comparing the average electrophoresis profiles of the two tissues, each sample contained 7 cases. The proteins spots of nasal polyposis and nasal mucosa tissues were 1532 and 1617. A total of 1 065 proteins spots were matched between the two tissues average electrophoresis profiles. (2)Twenty dif-ferential expression protein spots were incised from sliver staining gel randomly and digested in gel by TPCK Trypsin. Sixteen PMF were obtained by MALDI-TOF-MS, and 11 differential expression proteins were identified. CONCLUSION: In this study, the well-resolved reproducible 2-DE map of human nasal polyposis and nasal mucosa were established. Certain differential proteins were identified.