2',4'-dihydroxychalcone induces ferroptosis through ERO1A/GPX4 regulatory axis in cholangiocarcinoma

Background: Cholangiocarcinoma (CCA) , an aggressive Cancer often detected late, carries a grim prognosis. 2',4'-Dihydroxychalcone (2',4'-DHC), a flavonoid monomer isolated from Empetrum nigrum, has demonstrated notable anti-tumor activity in multiple Cancer types. However, its therapeutic mechanism in cholangiocarcinoma remains poorly understood, especially regarding the regulation of Ferroptosis, a mechanism that has not yet been fully elucidated in this disease.

Purpose: This study reveals that 2',4'-DHC induces Ferroptosis through modulation of the ERO1A/GPX4 signaling axis, thereby suppressing the progression of cholangiocarcinoma. These findings provide novel mechanistic insights and propose a potential therapeutic strategy for this highly refractory malignancy.

Methods: Proliferation, migration, and epithelial-mesenchymal transition assays were employed to determine the effects of 2',4'-DHC and ERO1A on CCA cells. Electron microscopy and Ferroptosis markers confirmed the involvement of Ferroptosis. Molecular docking and molecular dynamics simulations established direct targeting between 2',4'-DHC and ERO1A. Ubiquitination assays validated Ferroptosis potentiation. The anti-tumor efficacy of 2',4'-DHC was assessed in murine xenograft models.

Results: In our study, we first demonstrated that 2',4'-DHC inhibited the proliferation, migration and EMT progression of CCA cells, by inducing Ferroptosis, Mechanistically, 2',4'-DHC targeted ERO1A, leading to GPX4 ubiquitination and subsequent proteasomal degradation. In vivo, 2',4'-DHC suppressed tumor growth and significantly enhanced the therapeutic efficacy of oxaliplatin, without causing evident toxicity to major organs.

Conclusion: 2',4'-DHC induces Ferroptosis in CCA cells through the ERO1A/GPX4 axis and inhibits malignant progression. 2',4'-DHC drug-adjuvant therapy may enhance the efficacy of current Anticancer treatments. It also holds promise as a prognostic molecular marker and therapeutic target in CCA.

2′; 4 ′-dihydroxychalcone; Cholangiocarcinoma; ERO1A; Ferroptosis; GPX4.