Basic Study
Copyright ©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastrointest Oncol. Jan 15, 2025; 17(1): 99376
Published online Jan 15, 2025. doi: 10.4251/wjgo.v17.i1.99376
BIBR1532 inhibits proliferation and metastasis of esophageal squamous cancer cells by inducing telomere dysregulation
Qin Wang, Qing-Rong Li, Lei Xu, Zi-Chun Yuan, Xiao Liu, Mao-Ju Tang, Man Luo, Xiao-Wu Zhong, Qiang Ma, Xiao-Lan Guo
Qin Wang, Qing-Rong Li, Zi-Chun Yuan, Man Luo, Xiao-Wu Zhong, Qiang Ma, Xiao-Lan Guo, Department of Clinical Laboratory, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China
Qin Wang, Qing-Rong Li, Lei Xu, Zi-Chun Yuan, Xiao Liu, Mao-Ju Tang, Man Luo, Xiao-Wu Zhong, Qiang Ma, Xiao-Lan Guo, School of Laboratory Medicine & Translational Medicine Research Center, North Sichuan Medical College, Nanchong 637000, Sichuan Province, China
Co-first authors: Qin Wang and Qing-Rong Li.
Co-corresponding authors: Qiang Ma and Xiao-Lan Guo.
Author contributions: Guo XL and Ma Q contributed to conceptualization; Yuan ZC, Liu X, and Zhong XW contributed to investigation; Wang Q, Li QR, Xu L, Yuan ZC, Liu X, Tang MJ, Luo M, and Zhong XW contributed to methodology; Wang Q, Li QR, and Ma Q contributed to writing-original draft; Guo XL contributed to writing-review and editing; and all authors have read and approved the manuscript.
Supported by the Scientific Research Development Plan Project or the Scientific Research Foundation for Advanced Talents, Affiliated Hospital of North Sichuan Medical College, No. 2023MPZK017, No. 2023ZD001, No. 2023-2ZD002, and No. 2023GC009; Science and Technology Support Program of Nanchong, No. 22SXQT0001; Youth Medical Innovation Research Project, or Medical Research Project of Sichuan Province, No. Q23047 and No. S23020; and Development of a Scientific Research Plan for the Doctoral Scientific Research Foundation of the North Sichuan Medical College, No. CBY22-ZDA03.
Institutional animal care and use committee statement: The studies involving animals were reviewed and approved by the Ethics Committee of the North Sichuan Medical College (approval No. 2023033).
Conflict-of-interest statement: The authors declared that there are no conflicts of interest.
Data sharing statement: The data supporting the findings of this study are available from the corresponding author upon reasonable request.
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: Xiao-Lan Guo, MD, PhD, Professor, Department of Clinical Laboratory, Affiliated Hospital of North Sichuan Medical College, No. 1 South Maoyuan Road, Shunqing District, Nanchong 637000, Sichuan Province, China. alan5200@hotmail.com
Received: July 21, 2024
Revised: October 5, 2024
Accepted: November 1, 2024
Published online: January 15, 2025
Processing time: 144 Days and 0.7 Hours
Abstract
BACKGROUND

Esophageal squamous cell carcinoma (ESCC) is a malignant tumor with high morbidity and mortality, and easy to develop resistance to chemotherapeutic agents. Telomeres are DNA-protein complexes located at the termini of chromosomes in eukaryotic cells, which are unreplaceable in maintaining the stability and integrity of genome. Telomerase, an RNA-dependent DNA polymerase, play vital role in telomere length maintain, targeting telomerase is a promising therapeutic strategy for cancer.

AIM

To investigate the efficacy and underlying mechanisms of BIBR1532, a telomerase inhibitor, in ESCC.

METHODS

KYSE150 and KYSE410 cells were cultured and exposed to various concentrations of BIBR1532. Cell viability was assessed at 48 hours and 72 hours to determine the IC50 values. The effects of BIBR1532 on ESCC cell proliferation, migration, and cellular senescence were evaluated using the cell counting kit-8 assay, plate colony formation assay, scratch assay, transwell assay, and β-galactosidase staining, respectively. Western blotting was performed to detect the expression of proteins in BIBR1532-treated ESCC cells, such as human telomerase reverse transcriptase (hTERT), key molecules involved in DNA damage response (DDR) or cellular senescence, as well as telomere-binding proteins. Additionally, a tumor-bearing nude mouse model was established to evaluate the anti-cancer effect of BIBR1532 in vivo.

RESULTS

The IC50 values for KYSE150 and KYSE410 cells after 48 hours of BIBR1532 exposure were 48.53 μM and 39.59 μM, respectively. These values decreased to 37.22 μM and 22.71 μM, respectively, following a longer exposure of 72 hours. BIBR1532 exhibited dose-dependent effects on KYSE150 and KYSE410 cells, including decreased hTERT expression, inhibition of proliferation and metastasis, and induction of cellular senescence. Mechanistically, BIBR1532 upregulated the expression of the DDR protein, γ-H2AX, and activated the ataxia telangiectasia and Rad3-related protein (ATR)/ check point kinase 1 (CHK-1) and ataxia-telangiectasia mutated gene (ATM)/CHK2 pathways. BIBR1532 downregulated the expression of telomere-binding proteins, including telomeric-repeat binding factor 1 (TRF1), TRF2, protection of telomeres 1, and TIN2-interacting protein 1. In a nude mouse xenograft model, BIBR1532 significantly suppressed tumor growth, reduced hTERT expression, and increased γ-H2AX protein levels. Hematoxylin and eosin staining of various organs, including the heart, liver, spleen, lungs, and kidneys, revealed no apparent adverse effects.

CONCLUSION

BIBR1532 exerts anti-cancer effects on ESCC by inducing DDR through the ATR/CHK1 and ATM/CHK2 pathways and downregulating the expression of telomere-binding proteins.

Keywords: Esophageal squamous cell carcinoma; BIBR1532; Human telomerase reverse transcriptase; DNA damage response; Telomere-binding proteins

Core Tip: BIBR1532 inhibited proliferation and metastasis of esophageal squamous cell carcinoma (ESCC) cells in a dose-dependent manner, which also effectively blocked the growth of ESCC in tumor-bearing nude mouse model. Mechanistically, BIBR1532 downregulated the expression of telomere-binding proteins, upregulated the expression of phosphorylated histone H2AX, ataxia telangiectasia and Rad3-related protein/check point kinase 1 (CHK-1) and ataxia-telangiectasia mutated gene/CHK2 pathways. The in vitro and in vivo studies showed that BIBR1532 is a potential chemotherapeutic drug for ESCC.