Basic Study
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 14, 2022; 28(26): 3177-3200
Published online Jul 14, 2022. doi: 10.3748/wjg.v28.i26.3177
Involvement of Met receptor pathway in aggressive behavior of colorectal cancer cells induced by parathyroid hormone-related peptide
María Belén Novoa Díaz, Pedro Carriere, Graciela Gigola, Ariel Osvaldo Zwenger, Natalia Calvo, Claudia Gentili
María Belén Novoa Díaz, Pedro Carriere, Graciela Gigola, Natalia Calvo, Claudia Gentili, Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur (UNS)- INBIOSUR (CONICET-UNS), Bahía Blanca 8000, Buenos Aires, Argentina
Ariel Osvaldo Zwenger, Centro de Estudios Clínicos SAGA, CEC SAGA, Santiago de Chile 8320000, Chile
Author contributions: Novoa Díaz MB and Carriere P contributed to conceptualization, methodology, investigation, formal analysis, visualization, and manuscript drafting, review, and editing; Gigola G and Zwenger AO contributed to conceptualization, methodology, and investigation; Calvo N contributed to conceptualization, methodology, investigation, formal analysis, visualization, supervision, and manuscript drafting, review, and editing; Gentili C contributed to conceptualization, methodology, resources, investigation, formal analysis, visualization, supervision, manuscript drafting, review, and editing, project administration, and funding acquisition.
Supported by the Agencia Nacional de Promoción Científica y Tecnológica, No. PICT-2013-1441; Consejo Nacional de Investigaciones Científicas y Técnicas, No. PIP11220150100350; Instituto Nacional del Cáncer Asistencia Financiera II, RESOL 493/14, No. 2002-4395-14-1; Instituto Nacional del Cáncer Asistencia Financiera III-2016-2017, RESOL-2016-1006-E-APN-MS, No. 2002-3862-16-1 CANCER; Universidad Nacional del Sur, No. PGI: 24/B230 and No. PGI: 24/B303; and Fundación Alberto J. Roemmers of Argentina.
Institutional review board statement: The study was reviewed and approved by the committee of experts on the subject conformed by members of the Department of Biology, Biochemistry and Pharmacy of the National University of the South.
Institutional animal care and use committee statement: All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All experiments with animals were approved by a local animal committee for ethics (CICUAE-UNS, institutional endorsement updated to 2021), and were carried out in accordance with the National Institutes of Health guide for the care and use of Laboratory animals (NIH 1996).
Conflict-of-interest statement: There are no conflicts of interest to report.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE Guidelines, and the manuscript was prepared and revised according to the ARRIVE Guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (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: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Claudia Gentili, PhD, Professor, Research Scientist, Departamento de Biología, Bioquímica y Farmacia, Universidad Nacional del Sur (UNS)- INBIOSUR (CONICET-UNS), 670 San Juan, Bahía Blanca 8000, Buenos Aires, Argentina. cgentili@criba.edu.ar
Received: January 13, 2022
Peer-review started: January 13, 2022
First decision: March 8, 2022
Revised: March 21, 2022
Accepted: May 27, 2022
Article in press: May 27, 2022
Published online: July 14, 2022
Processing time: 180 Days and 17.1 Hours
ARTICLE HIGHLIGHTS
Research background

Colorectal cancer (CRC) patients usually relapse and die due to the development of metastasis and resistance to chemotherapeutic drugs employed in the treatment. Parathyroid hormone-related peptide (PTHrP) is a cytokine that has a key role in the carcinogenesis and the progression of several tumors. Our research group found that the binding of PTHrP to its receptor, the parathyroid hormone receptor type 1 (PTHR1), promotes events associated with the aggressive phenotype of colorectal cancer cells. The receptor tyrosine kinase Met is associated with metastasis and chemoresistance in CRC.

Research motivation

The mechanisms that lead to an aggressive tumor behavior and a worse response to CRC treatment are still unknown. To date, it has not been studied whether there is a cross-talk between the PTHrP and Met signaling pathways to trigger molecular mechanisms associated with CRC progression.

Research objectives

To investigate the relationship between PTHR1, PTHrP, and Met in the aggressive behavior of CRC cells. These findings could have implications for the future diagnosis, prognosis, and treatment of patients with CRC.

Research methods

By RT-PCR and Western blot analysis, we studied whether PTHrP modulates the expression of Met and its activation in CRC cells. By Western blot and using specific inhibitors, we also evaluated the signaling pathways involved in Met activation induced by PTHrP. By Trypan blue technique, Wound healing assay, Western blot, and morphological analysis, we evaluated whether the Met signaling pathway mediates the effects induced by PTHrP in HCT116 cells. By the immunohistochemistry technique, we evaluated the expression of Met and the PTHrP receptor, PTHR1, in HCT116 cell xenografts of nude mice in response to the treatment with or without PTHrP. An observational analysis of human samples was performed to validate the findings obtained by in vitro and in vivo assays.

Research results

PTHrP modulated the expression of Met and promoted its activation through Src kinase and the mitogen-activated protein kinase (MAPK) signaling pathway in human HCT116 cells. The Met signaling pathway triggered by PTHrP participated in cellular events related to the aggressive behavior of human HCT116 cells such as proliferation, morphological changes associated with epithelial to mesenchymal transition, and migration and in the decrease of the sensitivity to chemotherapy drugs. PTHrP increased the protein expression of its receptor, PTHR1, and of Met in HCT116 cell tumor xenografts. PTHR1 was highly expressed in well-differentiated human CRC samples. In contrast, Met showed an expression pattern that was increased in poorly differentiated CRC tumors.

Research conclusions

PTHrP promotes events associated with a worse behavior of CRC cells through Met activation. To date, our in vitro observations suggest that the binding of PTHrP to its receptor, PTHR1, promotes the regulation of Met gene expression and also its activation through the Src kinase and the MAPKs pathway. Once activated, Met signaling leads to molecular changes in the tumor cell to promote events associated with the aggressive behavior of CRC cells. Our previous data together with the findings shown herein reveal that PTHrP in vivo modulates the expression of markers linked to tumor progression (including Met) and also of its own receptor. These events could favor the tumor phenotype in the CRC animal model studied in this work.

Research perspectives

In the future, it will be of great interest to evaluate whether Met mediates other molecular mechanisms and events triggered by PTHrP that are involved in CRC progression. Concerning translational relevance, more studies are needed in the areas of basic and clinical oncology.