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©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
Novel therapeutic diiminoquinone exhibits anticancer effects on human colorectal cancer cells in two-dimensional and three-dimensional in vitro models
Alissar Monzer, Kevork Wakimian, Farah Ballout, Samar Al Bitar, Amani Yehya, Mariam Kanso, Nour Saheb, Ayman Tawil, Samer Doughan, Maher Hussein, Deborah Mukherji, Walid Faraj, Hala Gali-Muhtasib, Wassim Abou-Kheir
Alissar Monzer, Farah Ballout, Samar Al Bitar, Department of Biology, American University of Beirut, Beirut 1107-2020, Lebanon
Kevork Wakimian, Amani Yehya, Wassim Abou-Kheir, Department of Anatomy, Cell Biology and Physiological Sciences, American University of Beirut, Beirut 1107-2020, Lebanon
Mariam Kanso, Samer Doughan, Maher Hussein, Walid Faraj, Department of Surgery, American University of Beirut Medical Center, Beirut 1107-2020, Lebanon
Nour Saheb, Ayman Tawil, Department of Pathology and Laboratory Medicine, American University of Beirut Medical Center, Beirut 1107-2020, Lebanon
Deborah Mukherji, Department of Internal Medicine, American University of Beirut Medical Center, Beirut 1107-2020, Lebanon
Hala Gali-Muhtasib, Department of Biology and Center for Drug Discovery, American University of Beirut, Beirut 1107-2020, Lebanon
Author contributions: Monzer A carried out lab work as part of her PhD thesis, wrote the manuscript, and performed data analysis and interpretation of data (e.g., biostatistics, statistical analysis, and editing); Wakimian K, Ballout F, Al Bitar S, and Yehya A performed initial lab work and participated in data collection; Faraj W, Tawil A, Doughan S, Hussein M, Kanso M, and Saheb N helped in clinical data curation and the consent form for colon cancer patient samples at the American University of Beirut Medical Center; Gali-Muhtasib H and Abou-Kheir W conceived the project, supervised the work, and edited the manuscript draft; All authors have reviewed and approved the final manuscript.
Institutional review board statement: All specimens from the patients were obtained after their informed consent. All experiments involving human subjects were performed in agreement with all ethical considerations of the Institutional Review Board.
Institutional animal care and use committee statement: Prior to any mouse experiment, all mice protocols were reviewed and approved by the Institutional Animal Care and Use Committee (American University of Beirut, Institutional Animal Care and Use Committee).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The ARRIVE Guidelines have been adopted. 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: Wassim Abou-Kheir, MSc, PhD, Associate Professor, Department of Anatomy, Cell Biology and Physiological Sciences, American University of Beirut, Bliss street-Hamra, Beirut 1107-2020, Lebanon.
wa12@aub.edu.lb
Received: March 18, 2022
Peer-review started: March 18, 2022
First decision: June 11, 2022
Revised: June 24, 2022
Accepted: August 5, 2022
Article in press: August 5, 2022
Published online: September 7, 2022
Processing time: 165 Days and 12.7 Hours
ARTICLE HIGHLIGHTS
Research background
Colorectal cancer (CRC) is a multistep genetic disorder caused by sequential mutational events in signal transduction pathways occurring along with progression of the cancer. Quinone containing compounds have been reported as one of the promising novel anticancer therapeutics against CRC. However, the effects of diiminoquinone (DIQ) on CRC stem cells have not been extensively investigated yet.
Research motivation
To explore the promising anticancer effects of a novel quinone, DIQ, on CRC.
Research objectives
To investigate the anticancer potential of novel therapeutic DIQ on CRC using two-dimensional and three-dimensional models.
Research methods
MTT and trypan blue exclusion assays were employed to assess the anti-proliferative effect of DIQ on HCT116 and HT29 cell lines in vitro. Propidium iodide DNA and dihydroethidium staining were performed to determine cell cycle distribution and reactive oxygen species production in response to DIQ, respectively. Wound healing and transwell invasion assays were used to determine the invasion and migration ability of DIQ, respectively. Then, a sphere formation model was used to evaluate the potency of DIQ on targeting cancer stem cells in CRC cells for up to five generations. Immunofluorescent analysis and western blot were performed to elucidate the mechanism of action of DIQ in CRC. Organoid model was used to assess DIQ response on established organoids from fresh colorectal tissue samples from consenting patients.
Research results
DIQ reduced the self-renewal capacity of CRC cells and targeted the growth of colon cancer patient-derived organoids. DIQ downregulated the expression of key markers involved in the oncogenic stem cell Wnt/-catenin, AKT, and ERK signaling pathways that are involved in CRC tumorigenesis. Also, DIQ decreased proliferation, migration, and invasion and induced apoptosis, cell-cycle arrest, and reactive oxygen species.
Research conclusions
Our findings strongly suggest that DIQ could be a promising novel therapeutic for the treatment of CRC patients. This study represents the first documentation of the molecular mechanism of the novel anticancer therapeutics DIQ via targeting cancer stem cells, findings that have potential therapeutic implications for colon cancer patients.
Research perspectives
Further research on the DIQ mechanisms that are involved in CRC tumorigenesis is needed to be performed in the future. A larger cohort is still required to further investigate and evaluate the effects of DIQ in translational medicine.
