中国输血杂志2025，Vol.38Issue(8)：999-1007,9.DOI:10.13303/j.cjbt.issn.1004-549x.2025.08.001
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
8-methoxsalen photochemistry enhances tumor cell immunogenicity by inducing ferroptosis in B16 cells
摘要
Abstract
Objective To study the efficacy of photosensitizer 8-methoxsalen(8-MOP)combined with ultraviolet radia-tion 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/cm2),and then cultured in a constant temperature incubator(37℃,5％CO2)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 oxida-tive damage of tumor cells;Changes of Fe2+,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 de-tect 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 immunogenic-ity of tumor cells.5)1×105 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 immedi-ately 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,Fe2+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 immunoge-nicity of tumor cells,release more tumor antigens,promote the maturation of DC,present antigens better,thereby facilita-ting subsequent immune activation.关键词
铁死亡/免疫原性/抗原递呈/黑色素瘤/体外光化学疗法Key words
ferroptosis/immunogenicity/antigen presentation/melanoma/extracorporeal photochemotherapy分类
医药卫生引用本文复制引用
钟妍,林雨薇,陈伟,田力,李玲,刘忠..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[J].中国输血杂志,2025,38(8):999-1007,9.基金项目
中国医学科学院医学与健康科技创新工程(2021-1-I2M-060) （2021-1-I2M-060）
