富马酸二甲酯对鼻咽癌细胞辐射敏感性的影响及作用机制  

Effect and Mechanism of Dimethyl Fumarate on Radiosensitivity of Nasopharyngeal Carcinoma Cells

作　　者：王凯 韩琳[2] 王岳衡[1] WANG Kai;HAN Lin;WANG Yueheng(Hepingli Hospital,Beijing,China 100013;Beijing Puren Hospital,Beijing,China 100062)

机构地区：[1]北京市和平里医院,北京100013 [2]北京市普仁医院,北京100062

出　　处：《中国药业》2025年第7期36-44,共9页China Pharmaceuticals

摘　　要：目的探讨富马酸二甲酯(DMF)对人鼻咽癌细胞5-8F(简称5-8F细胞)辐射敏感性的影响及作用机制。方法选取北京市和平里医院耳鼻咽喉科2020年1月至2022年12月收治的62例鼻咽癌患者(之前未接受任何鼻咽癌相关治疗)的组织标本和核磁共振图像,按辐射敏感性的不同分为辐射敏感组(40例)和辐射抵抗组(22例)。通过四甲基偶氮唑蓝(MTT)法、克隆形成实验、流式细胞术评价DMF对5-8F细胞的活性、增殖、细胞周期和细胞凋亡的作用;采用免疫荧光染色法和免疫印迹(Western blot)法检测细胞核和细胞质中核因子-κB亚基p65(NF-κB p65)及上皮间质转换(EMT)标志物上皮钙黏素(E-cadherin)、神经钙黏素(N-cadherin)、波形蛋白(Vimentin)的蛋白表达水平。将18只小鼠随机分为Control组(不予IR和DMF处理)、电离辐射(IR)组、IR+DMF组,各6只。所有小鼠均复制鼻咽癌皮下移植瘤模型,考察DMF在小鼠体内对鼻咽癌辐射敏感性的影响。结果放疗2个月后,辐射敏感组患者的肿瘤消退明显,且组织中NF-kB p65和p-NF-kB p65均呈低表达;辐射抵抗组患者的肿瘤消退缓慢,NF-kB p65和p-NF-kB p65均呈高表达。DMF以浓度和时间依赖方式抑制5-8F细胞的活性,当DMF浓度为50.0μmol/L时,处理24 h对细胞活性的抑制率小于10%,而处理48,72 h时的细胞活性显著降低(74.21%,54.81%,P<0.05),故选取50.0μmol/L DMF处理24 h用于后续实验。与IR组比较,IR+DMF组5-8F细胞的克隆形成数量及存活分数均显著降低(P<0.05),磷酸化组蛋白(γ-H2AX)灶点数均显著增加(P<0.05),G2/M期细胞分布百分比和细胞凋亡率均显著升高(P<0.05),细胞核和细胞质中NF-κB p65及N-cadherin,Vimentin的蛋白表达水平均显著降低(P<0.05),E-cadherin的蛋白表达水平显著升高(P<0.05)。在鼻咽癌皮下移植瘤模型小鼠中,与IR组比较,IR+DMF组小鼠给药第1,2,3周后的肿瘤体积和肿瘤质量均显著减少(P<0.05),细胞核和细胞质中NF-κB p65�Objective To investigate the effect and mechanism of dimethyl fumarate(DMF)on the radiosensitivity of human nasopharyngeal carcinoma cell line 5-8F(referred to as 5-8F cells).Methods Tissue specimens and magnetic resonance imaging(MRI)images of 62 patients with nasopharyngeal carcinoma(who had not received any nasopharyngeal carcinoma-related treatment before)admitted to the Otolaryngology Department of Hepingli Hospital from January 2020 to December 2022 were selected and divided into the radiosensitive group(40 cases)and the radioresistant group(22 cases)according to their different radiosensitivity.MTT,colony formation and flow cytometry assay were conducted to evaluate the effects of DMF on cell viability,proliferation,cell cycle,and apoptosis of 5-8F cells.Immunofluorescence staining and Western blot were used to detect the expression levels of nuclear factor-κB subunit p65(NF-κB p65)protein in the nucleus and cytoplasm,and the epithelial-mesenchymal transition(EMT)markers such as E-cadherin,N-cadherin,and Vimentin protein.Eighteen mice were randomly divided into the control group(without IR or DMF treatment),ionizing radiation(IR)group,and IR+DMF group,with six mice in each group.All mice were replicated with nasopharyngeal carcinoma subcutaneous transplantation tumor models to investigate the effect of DMF on the radiosensitivity of nasopharyngeal carcinoma in mice.Results After two months of radiotherapy,patients in the radiosensitive group showed significant tumor regression and low-expressions of NF-κB p65 and p-NF-κB p65,while patients in the radioresistant group showed slow tumor regression and high-levels of NF-κB p65 and p-NF-κB p65.DMF inhibited the activity of 5-8F cells in a concentration-time-dependent manner,the inhibition rate of 50µmol/L DMF for 24 h was lower than 10%,and the cell viability decreased sharply at 48 and 72 h of treatment(74.21%,54.81%,P<0.05),so 50.0µmol/L DMF treatment for 24 h was selected for subsequent experiments.Compared with those in the IR group,the clone forma

关 键 词：富马酸二甲酯 人鼻咽癌细胞5-8F 辐射敏感性 细胞周期 细胞凋亡 核因子-ΚB 上皮间质转换 作用机制 

分 类 号：R969[医药卫生—药理学] R979.1[医药卫生—药学]