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©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jan 28, 2019; 25(4): 433-446
Published online Jan 28, 2019. doi: 10.3748/wjg.v25.i4.433
Interplay between post-translational cyclooxygenase-2 modifications and the metabolic and proteomic profile in a colorectal cancer cohort
Patricia Prieto, Rafael I Jaén, Daniel Calle, María Gómez-Serrano, Estefanía Núñez, María Fernández-Velasco, Paloma Martín-Sanz, Sergio Alonso, Jesús Vázquez, Sebastián Cerdán, Miguel Ángel Peinado, Lisardo Boscá
Patricia Prieto, Rafael I Jaén, Paloma Martín-Sanz, Sebastián Cerdán, Lisardo Boscá, Department of Metabolism and Physiopathology of Inflammatory Diseases, Instituto de Investigaciones Biomédicas Alberto Sols (CSIC-UAM), Madrid 28029, Spain
Patricia Prieto, Rafael I Jaén, María Gómez-Serrano, Estefanía Núñez, María Fernández-Velasco, Paloma Martín-Sanz, Jesús Vázquez, Lisardo Boscá, Centro de Investigación Biomédica en Red de Enfermedades Cardiovasculares (Ciber-CV), Instituto de Salud Carlos III (ISCIII), Madrid 28029, Spain
Daniel Calle, Laboratorio de Imagen Médica, Hospital Universitario Gregorio Marañón, Madrid 28007, Spain
María Gómez-Serrano, Estefanía Núñez, Jesús Vázquez, Laboratorio de Proteómica Cardiovascular, Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid 28029, Spain
María Fernández-Velasco, Instituto de Investigación Sanitaria del Hospital Universitario la Paz (IdiPaz), Madrid 28046, Spain
Sergio Alonso, Miguel Ángel Peinado, Programa de Medicina Predictiva y Personalizada del Cáncer (PMPPC), Fundación Instituto de investigación en ciencias de la salud Germans Trias i Pujol, Ctra Can Ruti, Badalona 08916, Spain
Author contributions: Prieto P and Jaén RI are the co-first authors. Prieto P performed the experiments, analyzed data and wrote the manuscript; Jaén RI performed the experiments and analyzed data, Calle D and Cerdán S designed, performed and analyzed the high resolution magic angle spinning experiments; Gómez-Serrano M, Núñez E, and Vázquez J designed and performed the proteomics analysis; Fernández-Velasco M and Martín-Sanz P revised critically the manuscript; Alonso S and Peinado MÁ collected colon biopsies from CRC patients and Boscá L conceived the experiments and revised critically the manuscript.
Supported by MINECO, No. SAF2017-82436R, SAF2016-75004R, RTC-2017-6283-1, PRB3 (IPT17/0019-ISCIII-SGEFI/ERDF) and BIO2015-67580P; Comunidad de Madrid, No. S2017/BMD-3686; Fundación Ramón Areces, No. 2016/CIVP18A3864; Instituto de Salud Carlos III, Spain, CIBERCV, No. CB/11/00222 and CB16/11/00277; FEDER, CIBEREHD; the Ministerio de Ciencia, Innovación y Universidades (MCNU); the Pro CNIC Foundation; and Severo Ochoa Center of Excellence, No. SEV-2015-0505.
Institutional review board statement: Colorectal cancer biopsies were obtained from 45 patients under informed consent, and were used in accordance with the procedures approved by Clinical Investigation ethics committees of the Germans Trias i Pujol Hospital (Badalona, Spain) and Bellvitge Hospital (Barcelona, Spain).
Conflict-of-interest statement: MAP is cofounder and equity holder of Aniling, a biotech company with no interests in this work. MAP lab has received research funding from Celgene. The rest of the authors declare no conflict of interest.
Data sharing statement: No additional data is available.
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/
Corresponding author: Lisardo Boscá, PhD, Professor, Department of Metabolism and Physiopathology of Inflammatory Diseases, Instituto de investigaciones Biomédicas “Alberto Sols”, Arturo Duperier 4, Madrid 28029, Spain. lbosca@iib.uam.es
Telephone: +34-91-4972747 Fax: +34-91-4972747
Received: October 17, 2018
Peer-review started: October 17, 2018
First decision: December 5, 2018
Revised: December 21, 2018
Accepted: January 9, 2019
Article in press: January 10, 2019
Published online: January 28, 2019
BACKGROUND
Colorectal cancer (CRC) is the second most common cause of cancer death worldwide. It is broadly described that cyclooxygenase-2 (COX-2) is mainly overexpressed in CRC but less is known regarding post-translational modifications of this enzyme that may regulate its activity, intracellular localization and stability. Since metabolic and proteomic profile analysis is essential for cancer prognosis and diagnosis, our hypothesis is that the analysis of correlations between these specific parameters and COX-2 state in tumors of a high number of CRC patients could be useful for the understanding of the basis of this cancer in humans.
AIM
To analyze COX-2 regulation in colorectal cancer and to perform a detailed analysis of their metabolic and proteomic profile.
METHODS
Biopsies from both healthy and pathological colorectal tissues were taken under informed consent from patients during standard colonoscopy procedure in the University Hospital of Bellvitge (Barcelona, Spain) and Germans Trias i Pujol University Hospital (Campus Can Ruti) (Barcelona, Spain). Western blot analysis was used to determine COX-2 levels. Deglycosylation assays were performed in both cells and tumor samples incubating each sample with peptide N-glycosidase F (PNGase F). Prostaglandin E2 (PGE2) levels were determined using a specific ELISA. 1H high resolution magic angle spinning (HRMAS) analysis was performed using a Bruker AVIII 500 MHz spectrometer and proteomic analysis was performed in a nano-liquid chromatography-tandem mass spectrometer (nano LC-MS/MS) using a QExactive HF orbitrap MS.
RESULTS
Our data show that COX-2 has a differential expression profile in tumor tissue of CRC patients vs the adjacent non-tumor area, which correspond to a glycosylated and less active state of the protein. This fact was associated to a lesser PGE2 production in tumors. These results were corroborated in vitro performing deglycosylation assays in HT29 cell line where COX-2 protein profile was modified after PNGase F incubation, showing higher PGE2 levels. Moreover, HRMAS analysis indicated that tumor tissue has altered metabolic features vs non-tumor counterparts, presenting increased levels of certain metabolites such as taurine and phosphocholine and lower levels of lactate. In proteomic experiments, we detected an enlarged number of proteins in tumors that are mainly implicated in basic biological functions like mitochondrial activity, DNA/RNA processing, vesicular trafficking, metabolism, cytoskeleton and splicing.
CONCLUSION
In our colorectal cancer cohort, tumor tissue presents a differential COX-2 expression pattern with lower enzymatic activity that can be related to an altered metabolic and proteomic profile.
Core tip: Although increased levels of cyclooxygenase-2 (COX-2) have been broadly related to colorectal cancer (CRC), high variability exists among patients. In our cohort, the electrophoretic band profile of COX-2 was altered in tumor tissue due to COX-2 glycosylation. Furthermore, prostaglandin E2 levels in tumor tissue were significantly lower than in normal colonic mucosa. In order to further characterize these samples, we have also detected an altered metabolic and proteomic profile in the tumor samples compared to controls that can be associated with the presence of glycosylated COX-2. These findings contribute to delineate the post-translational complexity and heterogeneity of molecular signaling regulation in CRC progression.
