亚甲基蓝介导的光动力治疗通过抑制SLC7A11表达诱导黑色素瘤细胞铁死亡  

Methylene-blue-mediated photodynamic therapy induces ferroptosis in melanoma cells by inhibiting SLC7A11 expression

作　　者：谭洁 蒋相康 张婉琪 杨桂红 鲁元刚 TAN Jie;JIANG Xiangkang;ZHANG Wanqi;YANG Guihong;LU Yuangang(Department of Plastic and Cosmetic Surgery,Daping Hospital,Army Medical University,Chongqing 400042,China;Department of Dermatology,Daping Hospital,Army Medical University,Chongqing 400042,China)

机构地区：[1]陆军军医大学大坪医院整形美容科,重庆400042 [2]陆军军医大学大坪医院皮肤科,重庆400042

出　　处：《免疫学杂志》2025年第1期29-35,43,共8页Immunological Journal

基　　金：国家自然科学基金(82272763)。

摘　　要：目的探索以亚甲基蓝(methylene blue,MB)为光敏剂的光动力治疗(photodynamic therapy,PDT)能否诱导黑色素瘤细胞铁死亡及其潜在机制。方法收集恶性黑色素瘤患者(5例)及恶性黑色素瘤淋巴结转移患者(5例)的病理石蜡切片,行普鲁士蓝染色、TUNEL染色、免疫组化染色,比较2组切片铁死亡程度;CCK8法、划痕实验分别检测不同处理条件下亚甲基蓝、亚甲基蓝介导的光动力治疗对B16F10的细胞毒性作用及细胞迁移能力的影响。Western blot检测B16F10细胞内铁死亡相关蛋白(GPX4、SLC7A11)的表达情况;酶标仪检测细胞内丙二醛、还原型/氧化型谷胱甘肽(GSH/GSSG)表达水平,流式细胞仪检测B16F10细胞内活性氧、Fe^(2+)、脂质过氧化物的表达水平。结果与转移至淋巴结的黑色素瘤细胞相比,未转移组铁死亡程度较高;MB、MB-PDT对B16F10的细胞毒性作用成剂量依赖性(P<0.0001);MB-PDT处理后,B16F10细胞的迁移能力被抑制(P<0.001);MB-PDT处理后,相比于对照组,细胞内GPX4、SLC7A11、GSH/GSSG表达水平降低(P<0.05),活性氧、Fe^(2+)、脂质过氧化物、丙二醛表达水平升高(P<0.05);用铁死亡抑制剂Ferrostatin-1预处理后可抑制MB-PDT诱导的B16F10细胞毒性和铁死亡(P<0.05)。结论MB-PDT可以通过抑制SLC7A11诱导铁死亡从而抑制黑色素瘤的进展。Objective This study aimed to investigate whether photodynamic therapy(PDT)with methylene blue(MB)as a photosensitizer can induce ferroptosis in melanoma cells and its potential mechanisms.Methods The paraffin sections of malignant melanoma patients(5 cases)and malignant melanoma patients with lymph node metastasis(5 cases)were collected.Prussian blue,TUNEL and immunohistochemical staining were performed to compare the degree of ferroptosis between the two groups.CCK-8 assay and scratch assay were used to detect the cytotoxic effect of methylene blue and methylene-blue-mediated photodynamic therapy on B16F10 under different treatment conditions and the effect of cell migration ability.The expression of ferroptosis-related proteins(glutathione peroxidase 4,GPX4、solute carrier family 7 member 11,SLC7A11)in B16F10 cells was detected by Western blot.The expression levels of intracellular malondialdehyde(MDA)and GSH/GSSG were detected by microplate reader.The expression levels of reactive oxygen species(ROS),Fe^(2+)and lipid peroxide(LPO)in B16F10 cells were detected by flow cytometry.Results Compared with melanoma cells transferred to lymph nodes,the non-metastatic group had a higher degree of ferroptosis and a lower expression level of SLC7A11.The cytotoxicity of MB and MB-PDT on B16F10 was dosedependent(P<0.0001).The results of scratch assay showed that MB-PDT inhibited the migration ability of B16F10 cells(P<0.001).Compared with the control group,after MB-PDT treatment,the expression levels of GPX4,SLC7A11 and GSH/GSSG were decreased(P<0.05),and the expression levels of ROS,Fe^(2+),LPO and MDA were increased(P<0.05)in B16F10 cells.Pretreatment with Ferrostatin-1,an inhibitor of ferroptosis,inhibited MB-PDT-induced cytotoxicity and ferroptosis in B16F10 cells(P<0.05).Conclusion MB-PDT can induce ferroptosis of melanoma cells by inhibiting SLC7A11 expression,thereby inhibiting the progression of melanoma.

关 键 词：黑色素瘤 光动力治疗 铁死亡 

分 类 号：R739.5[医药卫生—肿瘤]