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

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: , 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.