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  3. 5-Aminolevulinic Acid-Mediated Photodynamic Therapy Induces Ferroptosis in Oral Leukoplakia and Oral Squamous Cell Carcinoma

5-Aminolevulinic Acid-Mediated Photodynamic Therapy Induces Ferroptosis in Oral Leukoplakia and Oral Squamous Cell Carcinoma

  • Antioxidants (Basel). 2026 Jan 26;15(2):167. doi: 10.3390/antiox15020167.
Lei Zhang 1,  Ying Han 1,  Qianyun Guo 1,  Xinyi Ni 1,  Hongwei Liu 1
Affiliations

Affiliation

  • 1 Department of Oral Medicine, Peking University School and Hospital of Stomatology, National Center of Stomatology, National Clinical Research Center for Oral Diseases, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Research Center of Engineering and Technology for Computerized Dentistry Ministry of Health, NMPA Key Laboratory for Dental Materials, Beijing 100081, China.
PMID: 41750549 DOI: 10.3390/antiox15020167
Abstract

5-Aminolevulinic acid (ALA)-mediated photodynamic therapy (PDT) is one of the treatment modalities for oral leukoplakia (OLK) and oral squamous cell carcinoma (OSCC). However, the role of Ferroptosis in ALA-PDT for OLK and OSCC remains unclear. Therefore, this study aimed to investigate whether ALA-PDT can induce Ferroptosis in OLK and OSCC. We detected relative cellular dehydrogenase activity (CCK-8 assay), long-term proliferative viability, Reactive Oxygen Species (ROS) generation, glutathione levels, and mitochondrial morphology after ALA-PDT. The expression of ferroptosis-related proteins was detected using Western blot. A tongue OSCC model was established in male BalB/c nude mice, and then ALA-PDT was performed. Immunohistochemical staining of Ki67, GPX4 and FTH1 was conducted to evaluate the effect of ALA-PDT. Subsequently, OLK and OSCC cells were pre-treated with ferrostatin-1 (Fer-1) before ALA-PDT. Relative cellular dehydrogenase activity, ROS generation, lipid peroxidation, Fe2+ levels, and ferroptosis-related protein expression were measured. Finally, OLK and OSCC cells were treated with a combination of ALA-PDT and erastin, and mitochondrial function was evaluated. In vitro study showed that ALA-PDT increased ROS generation and decreased GSH/GSSG ratio in OLK and OSCC cells. After ALA-PDT, mitochondrial morphology exhibited typical characteristics of Ferroptosis. In vivo experiments showed that immunohistochemistry (IHC) scores of Ki67, GPX4 and FTH1 in the tissues decreased after ALA-PDT. Moreover, pre-treatment with Fer-1 could reverse ROS levels, lipid peroxidation and intracellular Fe2+ accumulation in OLK and OSCC cells after ALA-PDT. Additionally, Fer-1 pre-treatment reversed the changes in protein expression induced by ALA-PDT. The combination of ALA-PDT and erastin significantly reduced mitochondrial O2•- production and decreased mitochondrial membrane potential. Above all, ALA-PDT can induce Ferroptosis in OLK and OSCC. The use of Ferroptosis agonists may enhance the therapeutic efficacy of ALA-PDT for OLK and OSCC.

Keywords

5-aminolevulinic acid; ferroptosis; oral leukoplakia; oral squamous cell carcinoma; photodynamic therapy.

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