Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Feb 14, 2015; 21(6): 1749-1758
Published online Feb 14, 2015. doi: 10.3748/wjg.v21.i6.1749
Immunohistochemical expression of SP-NK-1R-EGFR pathway and VDR in colonic inflammation and neoplasia
Raymond A Isidro, Myrella L Cruz, Angel A Isidro, Axel Baez, Axel Arroyo, William A González-Marqués, Carmen González-Keelan, Esther A Torres, Caroline B Appleyard
Raymond A Isidro, Myrella L Cruz, Angel A Isidro, Caroline B Appleyard, Department of Physiology and Pharmacology, Ponce School of Medicine and Health Sciences, Ponce, PR 00732, United States
Angel A Isidro, Axel Baez, Department of Pathology, Ponce School of Medicine and Health Sciences, Ponce, PR 00732, United States
Axel Arroyo, Department of Anatomy, Ponce School of Medicine and Health Sciences, Ponce, PR 00732, United States
William A González-Marqués, Carmen González-Keelan, Departments of Pathology and Laboratory Medicine, University of Puerto Rico School of Medicine, San Juan, PR 00936, United States
Esther A Torres, Department of Medicine, University of Puerto Rico School of Medicine, San Juan, PR 00936, United States
Author contributions: Isidro RA, Isidro AA, Arroyo A, Torres EA and Appleyard CB designed the research; Isidro RA, Cruz ML, Isidro AA and Appleyard CB performed the research; Baez A, Arroyo A, González-Marqués WA, González-Keelan C and Torres EA identified and provided tissue samples; Isidro RA, Cruz ML and Appleyard CB analyzed the data; Isidro RA drafted the manuscript; Cruz ML, Isidro AA, Baez A, Arroyo A, Gonzalez-Marqués WA, González-Keelan C, Torres EA and Appleyard CB critically revised manuscript for important intellectual content.
Supported by National Institutes of Health Grants, No. U56 CA126379 (to Isidro AA and Appleyard CB) and No. R25-GM082406 (to Isidro RA); and National Science Foundation: Puerto Rico Louis Stokes Alliance for Minority Participation, NSF Project No. 0601843 (to Isidro RA); The PSMHS Molecular Biology Core Lab, RCMI Grant No. RR003050/MD007579.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Caroline B Appleyard, PhD, Professor, Department of Physiology and Pharmacology, Ponce School of Medicine and Health Sciences, PO Box 7004, Ponce, PR 00732-7004, United States. cappleyard@psm.edu
Telephone: +1-787-8402575 Fax: +1-787-8411040
Received: August 6, 2014
Peer-review started: August 8, 2014
First decision: September 15, 2014
Revised: September 27, 2014
Accepted: November 18, 2014
Article in press: November 19, 2014
Published online: February 14, 2015
Processing time: 188 Days and 18 Hours
Abstract

AIM: To determine the expression of neurokinin-1 receptor (NK-1R), phosphorylated epidermal growth factor receptor (pEGFR), cyclooxygenase-2 (Cox-2), and vitamin D receptor (VDR) in normal, inflammatory bowel disease (IBD), and colorectal neoplasia tissues from Puerto Ricans.

METHODS: Tissues from patients with IBD, colitis-associated colorectal cancer (CAC), sporadic dysplasia, and sporadic colorectal cancer (CRC), as well as normal controls, were identified at several centers in Puerto Rico. Archival formalin-fixed, paraffin-embedded tissues were de-identified and processed by immunohistochemistry for NK-1R, pEGFR, Cox-2, and VDR. Pictures of representative areas of each tissues diagnosis were taken and scored by three observers using a 4-point scale that assessed intensity of staining. Tissues with CAC were further analyzed by photographing representative areas of IBD and the different grades of dysplasia, in addition to the areas of cancer, within each tissue. Differences in the average age between the five patient groups were assessed with one-way analysis of variance and Tukey-Kramer multiple comparisons test. The mean scores for normal tissues and tissues with IBD, dysplasia, CRC, and CAC were calculated and statistically compared using one-way analysis of variance and Dunnett’s multiple comparisons test. Correlations between protein expression patterns were analyzed with the Pearson’s product-moment correlation coefficient. Data are presented as mean ± SE.

RESULTS: On average, patients with IBD were younger (34.60 ± 5.81) than normal (63.20 ± 6.13, P < 0.01), sporadic dysplasia (68.80 ± 4.42, P < 0.01), sporadic cancer (74.80 ± 4.91, P < 0.001), and CAC (57.50 ± 5.11, P < 0.05) patients. NK-1R in cancer tissue (sporadic CRC, 1.73 ± 0.34; CAC, 1.57 ± 0.53) and sporadic dysplasia (2.00 ± 0.45) were higher than in normal tissues (0.73 ± 0.19). pEGFR was significantly increased in sporadic CRC (1.53 ± 0.43) and CAC (2.25 ± 0.47) when compared to normal tissue (0.07 ± 0.25, P < 0.05, P < 0.001, respectively). Cox-2 was significantly increased in sporadic colorectal cancer (2.20 ± 0.23 vs 0.80 ± 0.37 for normal tissues, P < 0.05). In comparison to normal (2.80 ± 0.13) and CAC (2.50 ± 0.33) tissues, VDR was significantly decreased in sporadic dysplasia (0.00 ± 0.00, P < 0.001 vs normal, P < 0.001 vs CAC) and sporadic CRC (0.47 ± 0.23, P < 0.001 vs normal, P < 0.001 vs CAC). VDR levels negatively correlated with NK-1R (r = -0.48) and pEGFR (r = -0.56) in normal, IBD, sporadic dysplasia and sporadic CRC tissue, but not in CAC.

CONCLUSION: Immunohistochemical NK-1R and pEGFR positivity with VDR negativity can be used to identify areas of sporadic colorectal neoplasia. VDR immunoreactivity can distinguish CAC from sporadic cancer.

Keywords: Colitis; Colon cancer; Dysplasia; Neurokinin; Vitamin D

Core tip: This study compares for the first time the expression of neurokinin-1 receptor, phosphorylated epidermal growth factor receptor, cyclooxygenase-2, and vitamin D receptor in samples from patients with sporadic colorectal neoplasia, colitis, and colitis-associated colorectal neoplasia and obtained from a Puerto Rican population. We believe that this knowledge could be of great use for the further identification of reliable markers of cancer risk and of potential therapeutic targets in a population where colorectal cancer is the deadliest cancer, and that our findings could be used in the day-to-day examination of colonic biopsies for establishing a diagnosis and distinguishing dysplasia from reactive inflammatory changes.