[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.


METHODS
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.


RESULTS
(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 silver 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.


CONCLUSIONS
In this study, the well-resolved reproducible 2-DE map of human nasal polyposis and nasal mucosa were established. Certain differential proteins were identified.
He
Guangxiang
G
Department of Otorhinolaryngology, Third Xiangya Hospital of Central South University, Changsha, 410013, China.
Sun
Hong
H
Wang
Tiansheng
T
Li
Cui
C
Liu
Huowang
H
D000328:Adult; D000368:Aged; D016022:Case-Control Studies; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009297:Nasal Mucosa / Q000378:metabolism* / Q000473:pathology; D009298:Nasal Polyps / Q000378:metabolism* / Q000473:pathology; D040901:Proteomics