Berberine suppresses colorectal cancer progression by inducing ferroptosis-mediated energy metabolism disorders

Qiang Sun; Kerong Tu; Qiqi Xu; Li Yan; Shangqin Yang; Jingxuan Wang; Liangliang Lv; Hongmei Liu; Lulu Cai

doi:10.1016/j.jare.2025.10.025

Berberine suppresses colorectal cancer progression by inducing ferroptosis-mediated energy metabolism disorders

J Adv Res. 2025 Oct 24:S2090-1232(25)00820-3. doi: 10.1016/j.jare.2025.10.025. Online ahead of print.

Authors

Qiang Sun¹, Kerong Tu¹, Qiqi Xu¹, Li Yan², Shangqin Yang³, Jingxuan Wang¹, Liangliang Lv³, Hongmei Liu¹, Lulu Cai⁴

Affiliations

¹ Personalized Drug Research and Therapy Key Laboratory of Sichuan Province, Department of Pharmacy, Sichuan Provincial Peoples Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu 610072, China.
² Personalized Drug Research and Therapy Key Laboratory of Sichuan Province, Department of Pharmacy, Sichuan Provincial Peoples Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu 610072, China; Department of Gastrointestinal Surgery, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu 610072, China.
³ School of Pharmacy, Southwest Medical University, Luzhou 646000, China.
⁴ Personalized Drug Research and Therapy Key Laboratory of Sichuan Province, Department of Pharmacy, Sichuan Provincial Peoples Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu 610072, China. Electronic address: cailulu@med.uestc.edu.cn.

PMID: 41139018
DOI: 10.1016/j.jare.2025.10.025

Abstract

Introduction: Berberine (BBR), the predominant isoquinoline alkaloid in Coptidis Rhizoma, exhibits remarkable anti- colorectal cancer (CRC) activity. However, whether BBR triggers CRC cell death through ferroptosis-associated disruption of energy metabolism remains to be elucidated.

Objective: To investigate if BBR induces mitochondrial energy metabolism disorder in CRC cells by regulating the ferroptosis signaling pathway.

Methods: BBR's effects on malignant phenotypes were evaluated in vitro (human cell line HCT116, murine cell line CT26 cells at 10, 20, 40 μM) and in vivo (80 mg/kg). Target engagement and mechanistic pathways were interrogated through RNA-sequence combined with convolutional neural network-based pathway prediction, corroborated by surface plasmon resonance, cellular thermal shift assay. Downstream validation mainly included quantification of Gli1, STAT3, GPX4, SLC7A11, and FTH1 expression via RT-qPCR, Western blot, immunofluorescence, and other molecular expression and functional confirmation experiments.

Results: BBR inhibited CRC cell proliferation with IC₅₀ value for HCT116 cells for 48 h at 19.86 ± 2.31 μM, and for CT26 cells for 48 h at 21.35 ± 2.63 μM. Concurrently, it elevated ferroptosis markers such as malondialdehyde, lactate dehydrogenase, Fe²⁺, and 4-hydroxynonenal, while suppressing ATP levels, superoxide dismutase activity, and energy metabolism-related enzymes. Graph convolutional network-based drug "on-target" pathway algorithm predicted Gli1 as top-9 target, and surface plasmon resonance confirmed direct BBR-Gli1 binding with KD value at 0.652 μM, cellular stability thermal assessment showed BBR stabilized Gli1 with thermal shift with ΔT = 2.3 °C. Mechanistically, BBR exerted its anti-CRC effects by inhibiting the Gli1/STAT3-ferroptosis negative regulation (Gli1/STAT3-FNR) axis, a novel regulatory pathway. Notably, BBR exhibited no significant organ or hematological toxicity in vivo at the experimental doses.

Conclusion: BBR triggers ferroptosis-mediated energy metabolism disorder by inhibiting Gli1/STAT3-FNR axis. This work provides a mechanistic support for BBR anti-CRC indications, and suggests an encouraging approach for treating CRC.

Keywords: Berberine; Colorectal cancer; Ferroptosis; Gli1/STAT3-FNR axis; Mitochondrial energy metabolism.