Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastrointest Oncol. May 15, 2023; 15(5): 810-827
Published online May 15, 2023. doi: 10.4251/wjgo.v15.i5.810
BZD9L1 benzimidazole analogue hampers colorectal tumor progression by impeding angiogenesis
Chern Ein Oon, Ayappa V Subramaniam, Lik Yang Ooi, Ashwaq Hamid Salem Yehya, Yeuan Ting Lee, Gurjeet Kaur, Sreenivasan Sasidharan, Beiying Qiu, Xiaomeng Wang
Chern Ein Oon, Ayappa V Subramaniam, Lik Yang Ooi, Yeuan Ting Lee, Gurjeet Kaur, Sreenivasan Sasidharan, Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang 11800, Malaysia
Ashwaq Hamid Salem Yehya, Cancer Research, Eman Biodiscoveries, Kedah 08000, Malaysia
Ashwaq Hamid Salem Yehya, Vatche and Tamar Division of Digestive Diseases, Department of Medicine, David Geffen School of Medicine at University of California Los Angeles, Los Angeles, CA 90095, United States
Beiying Qiu, Xiaomeng Wang, Academic Clinical Program, Duke-NUS Medical School, National University of Singapore, Singapore 169857, Singapore
Beiying Qiu, Singapore National Eye Centre, Singapore Eye Research Institute, Singapore 168751, Singapore
Xiaomeng Wang, Singapore National Eye Centre, Singapore Eye Research Institute, Singapore 169857, Singapore
Author contributions: Oon CE and Subramaniam AV remain to have equal contributions as co-first authors; Oon CE, Subramaniam AV, Ooi LY, and Qiu B carried out the experiments; Lee YT synthesized the BZD9L1 compound; Oon CE drafted and reviewed the manuscript; Subramaniam AV and Ooi LY provided scientific input for “Discussion”; Oon CE, Subramaniam AV, Ooi LY, Qiu B, and Kaur G analyzed the data; Sasidharan S provided scientific input for in vivo study; Wang X derived the ex vivo angiogenesis models; Oon CE conceived and designed the experiments, and supervised the project; and all the authors have read and approved the final manuscript.
Supported by the Ministry of Higher Education Malaysia for the Fundamental Research Grant Scheme, No. FRGS/1/2021/SKK06/USM/02/7.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the National University of Singapore Institutional Animal Care and Use Committee guidelines (approval No. 2020/SHS/1597) and Universiti Sains Malaysia Animal Ethical Committee [approval No. USM/IACUC/2017/(105)(872)].
Conflict-of-interest statement: The authors declare that they have no conflict of interest.
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:
Corresponding author: Chern Ein Oon, DPhil, PhD, Associate Professor, Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Gelugor, Penang 11800, Malaysia.
Received: January 3, 2023
Peer-review started: January 3, 2023
First decision: February 13, 2023
Revised: February 17, 2023
Accepted: April 21, 2023
Article in press: April 21, 2023
Published online: May 15, 2023
Research background

The growth and spread of colorectal cancer (CRC) are highly dependent on angiogenesis. Epigenetic regulation of the genes in endothelial cells (ECs) in the vicinity of tumor cells plays a vital role in tumor angiogenesis. Sirtuins are class III histone deacetylase enzymes that are implicated in angiogenesis. Their potential roles in cancer have stimulated investigation to seek potent and selective sirtuin (SIRT) inhibitors, potentially leading to new therapeutic breakthroughs. BZD9L1 is a reported small molecule inhibitor with anticancer activities. However, its potential as an anti-angiogenic agent has not been explored.

Research motivation

A patient’s prognosis and survival rate remain heterogenous for which tumor attributes, dynamic host response factors, and treatment quality may be accountable. Some CRC patients become resilient to these anti-angiogenic drugs and standard therapies such as chemotherapy and radiation. Hence, this work opens a new avenue for the establishment of a potential novel anti-angiogenic agent through sirtuin inhibition in tumor angiogenesis.

Research objectives

To determine the anti-angiogenic activity of BZD9L1 benzimidazole analogue in CRC.

Research methods

The in vitro experiments comprise cell viability, adhesion. spheroid sprouting, quantitative polymerase chain reaction (qPCR), angiogenesis protein array, cell cycle and apoptosis analyses via flow cytometry and indirect co-culture. Mouse choroids were employed to assess the negative impact of BZD9L1 on sprouting and vessel regression. HCT116 CRC cells were injected subcutaneously into athymic nude mice and treated with vehicle control or BZD9L1 at 50 mg/kg and 250 mg/kg. Hematoxylin and eosin staining was performed to determine the percentage of necrosis in the tumor section. Finally, immunohistochemistry and qPCR were conducted to investigate the expression of Ki67 protein and murine CD34/ CD31 as well as SIRT1 and SIRT2, respectively.

Research results

Findings from this study highlighted the ability of BZD9L1 to inhibit EC functions in in vitro, ex vivo and co-culture models. Additionally, BZD9L1 retarded tumor growth in vivo compared to the vehicle control group. Overall, the findings underscore the potential of BZD9L1 to treat CRC.

Research conclusions

BZD9L1 impeded angiogenesis in ECs, mouse choroid tissues and the CRC xenograft model. This study provides valuable insights into BZD9L1 as a potential anti-angiogenic agent in CRC.

Research perspectives

Findings from this study may provide the basis for BZD9L1 benzimidazole analogue as a targeted therapy for the treatment of CRC.