8-methoxsalen photochemistry enhances tumor cell immunogenicity by inducing ferroptosis in B16 cells
10.13303/j.cjbt.issn.1004-549x.2025.08.001
- VernacularTitle:8-甲氧基补骨脂素光化学通过诱导B16细胞铁死亡提高肿瘤细胞免疫原性
- Author:
Yan ZHONG
1
;
Yuwei LIN
1
;
Wei CHEN
1
;
Li TIAN
1
;
Ling LI
2
;
Zhong LIU
1
Author Information
1. Institute of Blood Transfusion, Chinese Academy of Medical Sciences& Peking Union Medical College, Chengdu 610052, China; Key Laboratory of Transfusion Adverse Reactions, Chinese Academy of Medical Sciences, Chengdu 610052, China
2. School of Clinical Medicine, Southwest Jiaotong University and The Third People's Hospital of Chengdu, Chengdu 610014, China
- Publication Type:Journal Article
- Keywords:
ferroptosis;
immunogenicity;
antigen presentation;
melanoma;
extracorporeal photochemotherapy
- From:
Chinese Journal of Blood Transfusion
2025;38(8):999-1007
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To study the efficacy of photosensitizer 8- methoxsalen (8-MOP) combined with ultraviolet radiation A (UVA) in inducing ferroptosis in mouse melanoma B16 cells, and to assess the resultant changes in immunogenicity, and their impact on subsequent immune activation after treatment. Methods: 1) Mouse melanoma B16 cells were cultured and treated with 8-MOP (100 ng/mL) and UVA (4 J/cm
), and then cultured in a constant temperature incubator (37℃, 5%CO
) for 24 hours after irradiation. 2) CCK8 (cell proliferation and toxicity) detection kit was used to detect the death rate of tumor cells. 3) LPO (lipid peroxide) and GSH (glutathione) detection kits were used to detect the degree of oxidative damage of tumor cells; Changes of Fe
, mitochondrial membrane potential (JC-1) and BODIPY 581/591 C11 (lipid peroxidation detection kit) in tumor cells were detected by confocal microscope. Western blotting (WB) was performed to detect GPX4, SLC7A11 and NCOA4 to confirm ferroptosis. 4) The expression of HMGB1 (high mobility group protein 1), ATP and CRT (calreticulin) in the supernatant of tumor cell culture was detected by ELISA kit to evaluate the immunogenicity of tumor cells. 5) 1×10
B16 cells were injected subcutaneously into the skin of the back and neck of mice at a dose of 100 μL to construct a mouse melanoma model. Spleen mononuclear cells of tumor-bearing mice were extracted and immediately co-cultured with irradiated tumor cells for 48 h. Changes of dendritic cell (DC) maturity were detected by MHC-II, CD11c, CD80 and CD83 flow cytometry. Results: After UVAP, the survival rate of B16 cells decreased significantly (61.39±6.823 vs 84.81±7.026 vs 100.0±3.996, P<0.000 1, P<0.01). UVAP effectively induced ferroptosis in B16 cells, characterized by increased LPO and C11-bodipy lipid peroxidation, GSH depletion, Fe
accumulation, mitochondrial membrane depolarization, decreased GPX4 and SLC7A11 protein expression, and increased NCOA4 expression, all in line with the trend of ferroptosis. UVAP also enhanced tumor cell immunogenicity, evidenced by elevated release of ATP, CRT, and HMGB1. The immunogenicity of B16 cells increased, the expressions of ATP, CRT and HMGB1 increased, and the DC maturity increased (CD80: 31.92±4.071 vs 19.77±3.177; CD83: 21.40±4.787 vs 12.19±1.487, P<0.001, P<0.01). Conclusion: The combined action of 8-MOP and UVA can induce ferroptosis in B16 tumor cells, enhance the immunogenicity of tumor cells, release more tumor antigens, promote the maturation of DC, present antigens better, thereby facilitating subsequent immune activation.
