Blockade of calcium-activated chloride channel ANO1 ameliorates ionizing radiation-induced intestinal injury

Sep 29, 2026

Radiation enteritis is a frequent complication of radiotherapy, and effective protective strategies remain limited. This study examines whether targeting anoctamin 1 (ANO1) can mitigate radiation-induced intestinal injury while simultaneously improving tumor radiosensitivity. ANO1 expression changes after irradiation were measured using qPCR and Western blot. Functional validation used ANO1 agonists/inhibitors in mice and 3D organoids, with ion levels and membrane potential assessed by fluorescent probes and mechanism analysis by additional protein assays.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages11

Reading Time28 min

AccessFree to download

Introduction

Radiation enteritis burden and clinical management

Objectives

Methods

Molecular and protein assays

Animal models and organoid validation

Ion measurements and mechanistic studies

Results

ANO1 blockade effects and radiosensitization

Mechanistic link to NKCC1 and Cl- regulation

Conclusion

Journal of Advanced Research 79 (2026) 433–443 Contents lists available at ScienceDirect Journal of Advanced Research journal [homepage: www. elsevier. com/locate/jare](homepage: www. elsevier. com/locate/jare) Blockade of calcium-activated chloride channel ANO1 ameliorates ionizing radiation-induced intestinal injury Yuying Guo a, Tong Yuan a, Yuna Wang a, Lei Xiab,⁎, Junling Zhang a,⁎, Saijun Fan a,⁎ a Institute of Radiation Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Tianjin 300192, China b Department of Cancer Center, The Second Afﬁliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Immunotherapy, Chongqing 401336, China h i g h l i g h t s • ANO1 inhibition protects against radiation-induced intestinal damage. • CaCCinh-A01 blocks calciumactivated chloride currents and reduces ANO1 expression. • NKCC1 plays vital role in the radioprotective effect of chloride channel blockade. • Chloride channel blockade inhibits membrane depolarization and DNA damage in irradiated cells. • CaCCinh-A01 enhances colorectal cancer radiosensitivity, offering dual therapeutic beneﬁts. g r a p h i c a l a b s t r a c t a r t i c l e i n f o Article history: Received 17 November 2024 Revised 7 April 2025 Accepted 7 April 2025 Available online 8 April 2025 Keywords: ANO1 CaCCinh-A01 Radiation-induced intestine injury Colorectal cancer a b s t r a c t Introduction: Radiation enteritis is one of the most frequent clinical complications of radiotherapy (RT), yet few effective strategies currently exist to protect against that. Anoctamin 1 (ANO1) functions both asa chloride channel and a signal transduction protein, inﬂuencing numerous pathophysiological processes. Objectives: This study aimed to investigate whether targeting ANO1 could mitigate radiation-induced enteritis while enhancing tumor radiosensitivity. Methods: Quantitative PCR (qPCR) and Western blot (WB) were used to assess ANO1 expression and its changes after irradiation. Survival rates were recorded to evaluate the effects of ANO1 agonist and inhibitors. A cystic ﬁbrosis transmembrane conductance regulator (CFTR) inhibitor was administered to irradiated mice to investigate the role of chloride channel in radiation protection. qPCR and WB were executed to analyze the expression of relevant ion channels in intestinal epithelium. Functional validation was conducted using inhibitors in mice and 3D organoids. Fluorescent probe kits detected intracellular ion levels and membrane potential, and WB was performed to elucidate the underlying mechanisms. Finally, the radiosensitizing effect of CaCCinh-A01 was assessed in colorectal cancer (CRC) cells and validated in in vivo models. Results: Blocking the calcium-activated chloride channel (CaCC) protein ANO1, which is highly expressed in the colon, protects the intestine from radiation-induced damage. The ANO1 inhibitor CaCCinh-A01, suppresses CaCC currents, downregulates ANO1 protein expression, alleviates radiation-induced intestine injury, and enhances the radiosensitivity of CRC. Mechanistically, CaCCinh-A01 upregulates Na-KCl Cotransporter 1 (NKCC1) protein expression, leading to an increase in intracellular Cl- concentration ⁎ Corresponding authors. E-mail addresses: [xialei@cqmu.edu.cn](xialei@cqmu.edu.cn) (L. Xia), [zhangjunling@irm-cams.ac.cn](zhangjunling@irm-cams.ac.cn) (J. Zhang), [fansaijun@irm-cams.ac.cn](fansaijun@irm-cams.ac.cn) (S. Fan). [https://doi.org/10.1016/j.jare.2025.04.009](https://doi.org/10.1016/j.jare.2025.04.009) 2090-1232/© 2025 The Authors. Published by Elsevier B.V. on behalf of Cairo University. This is an open access article under the CC BY-NC-ND license ([http://creativecommons.org/licenses/by-nc-nd/4.0/](http://creativecommons.org/licenses/by-nc-nd/4.0/)). Y. Guo, T. Yuan, Y. Wang et al. Journal of Advanced Re

FAQs

What clinical problem does the study target?

The study targets radiation enteritis, a common complication of radiotherapy that lacks standardized, effective protective strategies.

How did researchers evaluate ANO1 involvement after irradiation?

They measured ANO1 expression changes using quantitative PCR and Western blot, and used inhibitors/agonists in mice and 3D organoids for functional validation.

What is the main finding regarding CaCCinh-A01?

CaCCinh-A01 blocks ANO1-associated calcium-activated chloride currents, reduces ANO1 expression, alleviates radiation-induced intestinal injury, and enhances colorectal cancer radiosensitivity.

What mechanism links chloride channel blockade to reduced DNA damage?

The study reports that CaCCinh-A01 increases NKCC1 expression, raises intracellular Cl- concentration, reduces membrane depolarization, and suppresses p53-mediated DNA damage signaling.

Sep 29, 2026

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