Sox-9 Immunohistochemistry Demonstrates Abnormal Distribution of Gastrointestinal Stem Cells in Specialized and Non-Specialized Barrett's Mucosa.
Mahmoud Goodarzi, Rujuta Kanhere, Lynda Corley, Arlene Correa, Jaffer Ajani, Stephen Swisher, Wayne Hofstetter, Asif Rashid, Dipen Maru. The University of Texas M.D.Anderson Cancer Center, Houston; Caris Research Institute, Caris Life Sciences, Newton
Background: Sox-9 is a gastrointestinal stem cell marker shown to be abnormally expressed in specialized Barrett's mucosa (BM)/intestinal metaplasia. The distribution of stem cell in cardia type mucosa in tubular esophagus (non-specialized BM) is unknown. We quantitatively assessed the stem cell distribution in two types of BM and oxyntic mucosa.
Design: Gastric body biopsies with unremarkable oxyntic mucosa,esophageal biopsies with non-specialized and specialized BM from patients with BM on endoscopy were analyzed. Sox-9 (rabbit antibody,Milipore,CA)immunohistochemistry was performed with an automated immunostainer (Leica Microsystems,IL). The SOX9 positive nuclei were quantitatively assessed by automated image analysis using Aperio's ScanScope XT (Aperio Technologies Inc,CA). Surface epithelium, upper half (body) and lower half (neck and base) of crypt were analyzed as separate overlays in the scanned images at 20X. Three compartments were marked to include only epithelial cells. An automated nuclear quantitation algorithm using the ScanScope was prepared based on color variations to achieve percent positive cells per lesion for each compartment. Mann–Whitney U test and SamplePower 2.0 software used for the statistical analysis.
Results: Twenty two biopsies from 17 adults were analyzed. Sox-9 nuclear staining was observed in lower crypt of the oxyntic mucosa with essentially no staining of the surface epithelium. In contrast, Sox-9 staining was seen in all three compartments of the non-specialized and specialized BM with florid staining in the multilayered columnar epithelium.
|Fundus (n=6)||Non-Specialized BM (n=8)||Specialized BM (n=8)||p value/Power|
|Total||17.9±9.3||59.6±24.3||66.2±29.3||0.001/0.95a, 0.001/0.96b, 0.72c|
|Surface||3.2±2.5||15.1±7.3||18.3±6.1||0.001/0.93a, 0.008/0.87b, 0.23c|
|Upper crypt||4.7±1.8||13.9±8.5||19.1±10.2||0.08/0.66a, 0.008/0.87b, 0.23c|
|Lower crypt||10.1±6.5||30.6±14||28.8±16.3||0.003/0.85 a, 0.013/0.68b, 0.72c|