Proteomic analysis of colonic crypts from normal, multiple intestinal neoplasia and p53-null mice: A comparison with colonic polyps

Citation
Ar. Cole et al., Proteomic analysis of colonic crypts from normal, multiple intestinal neoplasia and p53-null mice: A comparison with colonic polyps, ELECTROPHOR, 21(9), 2000, pp. 1772-1781
Citations number
59
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Chemistry & Analysis
Journal title
ELECTROPHORESIS
ISSN journal
0173-0835 → ACNP
Volume
21
Issue
9
Year of publication
2000
Pages
1772 - 1781
Database
ISI
SICI code
0173-0835(200005)21:9<1772:PAOCCF>2.0.ZU;2-7
Abstract
In order to observe cellular changes caused by mutation of the tumor suppre ssors, APC and p53, we have generated protein expression profiles of mouse colon epithelial cells using two-dimensional electrophoresis (2-DE). Crypts , polyps and stroma were isolated from normal, multiple intestinal neoplasi a (MIN) ana p53-null mice, each with a C57Black/6J background, and subjecte d to 2-DE in two separate pH ranges (pH 3-10 and pH 6-11). No significant d ifferences in protein expression patterns were observed between the normal, MIN and p53-null colon epithelial crypts. However, 64 proteins from the MI N polyps showed a 2-fold or greater difference in intensity that was statis tically significant as assessed by the Wilcoxon rank-sum test (p less than or equal to 0.05). Of these, calreticulin, carbonic anhydrase I and a new m ember of the glutathione-S-transferase theta family of proteins have so far been identified using an in-gel digestion protocol coupled with reversed-p hase high performance liquid chromatography (RP-HPLC) ion-trap mass spectro metry. In addition, 38 marker proteins have been identified in a continuing effort to generate a comprehensive 2-DE database of proteins expressed by mouse colon epithelial cells (these databases are available at http:/www. l udwig.edu.au/jpsl/jpSlhome.html.).