西达本胺联合姜黄素对皮肤T细胞淋巴瘤的作用及机制研究  

Antitumor effects and mechanisms of action of chidamide combined with curcumin in the treatment of cutaneous T-cell lymphoma

作　　者：王冠钰 孙佳辰 李婷婷 王艺萌[1] 张春雷[1] Wang Guanyu;Sun Jiachen;Li Tingting;Wang Yimeng;Zhang Chunlei(Department of Dermatology,Peking University Third Hospital,Beijing 100191,China)

机构地区：[1]北京大学第三医院皮肤科,北京100191

出　　处：《中华皮肤科杂志》2024年第8期728-738,共11页Chinese Journal of Dermatology

基　　金：国家自然科学基金(81972560);北京自然科学基金(7202231);北京大学医学部青年培育项目(BMU2020PYB023)。

摘　　要：目的研究西达本胺联合姜黄素对皮肤T细胞淋巴瘤(CTCL)的抗肿瘤作用及安全性。方法体外培养人CTCL细胞系HH和HuT 78,设置西达本胺(浓度0.4、0.8、1.6、3.2、6.4μmol/L)、姜黄素(1.25、2.5、5、10、20μmol/L)梯度浓度单用/联用,评估两者对HH、HuT 78细胞的联合用药指数(CI)。将细胞分为西达本胺组(0.4μmol/L西达本胺)、姜黄素组(10μmol/L姜黄素)、联合用药组(0.4μmol/L西达本胺+10μmol/L姜黄素)及溶剂对照组,培养48 h后采用MTS法检测细胞增殖,流式细胞仪检测细胞凋亡,实时定量PCR和Western印迹分别检测细胞凋亡相关基因核因子(NF)κB p65、B淋巴细胞瘤2(Bcl-2)和胱天蛋白酶3(caspase-3)mRNA和蛋白的表达。建立免疫缺陷小鼠HH细胞荷瘤模型,设西达本胺组(10 mg/kg西达本胺)、姜黄素组(100 mg/kg姜黄素)、联合用药组、溶剂对照组,连续灌胃12 d,测量给药后各组小鼠体重、肿瘤体积。第13天处死小鼠,取肿瘤组织通过原位末端转移酶标记法(TUNEL)染色检测肿瘤细胞的凋亡情况,定量PCR和Western印迹检测细胞凋亡相关基因和蛋白的表达。多组间差异采用单因素方差分析,组间两两比较采用LSD-t检验。结果0.4~6.4μmol/L西达本胺联合1.25~20μmol/L姜黄素对HH、HuT 78细胞的CI值均<1,二者联用显示协同作用。各组HuT 78、HH细胞培养48 h,联合用药组增殖率低于西达本胺组、姜黄素组(均P<0.05);联合用药组HuT 78细胞凋亡率(70.47%±7.87%)高于西达本胺组、姜黄素组及对照组(31.95%±9.43%、37.23%±10.74%、11.76%±5.65%,均P<0.001);联合用药组HH细胞凋亡率(28.31%±1.70%)高于西达本胺组、姜黄素组及对照组(21.29%±3.61%、18.74%±1.82%、3.18%±1.00%,均P<0.001);与对照组、西达本胺组及姜黄素组比较,联合用药组caspase-3 mRNA及剪切体蛋白的表达明显增加(均P<0.05),NF-κB p65、Bcl-2 mRNA及蛋白的表达明显降低(均P<0.05)。小鼠体内实验第13天时,联合用药组�Objective To evaluate the efficacy and safety of chidamide combined with curcumin in the treatment of cutaneous T-cell lymphoma(CTCL).Methods Human CTCL cell lines HH and HuT-78 were cultured in vitro and treated with gradient concentrations of chidamide(0.4,0.8,1.6,3.2,and 6.4μmol/L)and curcumin(1.25,2.5,5,10,and 20μmol/L)alone or in combination,and the combination index(CI)of chidamide and curcumin for HH and HuT-78 cells was evaluated.Cultured HH/HuT-78 cells were divided into chidamide group(treated with 0.4μmol/L chidamide),curcumin group(treated with 10μmol/L curcumin),combination group(treated with 0.4μmol/L chidamide+10μmol/L curcumin),and solvent control group(treated with dimethyl sulfoxide);after 48-hour treatment,the MTS assay was performed to evaluate the cell viability,flow cytometry to detect cell apoptosis and analyze cell cycle,and real-time quantitative PCR(RT-PCR)and Western blot analysis were conducted to determine the mRNA and protein expression of apoptosis-related genes nuclear factor(NF)-κB p65,B-cell lymphoma 2(Bcl-2),and caspase-3,respectively.A tumor-bearing mouse model was established with HH cells in immunodeficient mice.These tumor-bearing mice were randomly divided into 4 groups:chidamide group(gavaged with 10 mg/kg chidamide),curcumin group(gavaged with 100 mg/kg curcumin),combination group,and solvent control group.The treatment was administered daily for 12 days,and body weight and tumor size were measured.On day 13,these mice were sacrificed,and tumor tissues were collected.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining was performed to detect apoptosis of tumor cells,and RT-PCR and Western blot analysis were conducted to determine the expression of apoptosis-related genes and proteins.Differences among multiple groups were analyzed using one-way analysis of variance,and multiple comparisons were performed using least significant difference-t test.Results The CI values of chidamide(0.4-6.4μmol/L)combined with curcumin(1.25-20μmol/L)

关 键 词：淋巴瘤 T细胞 皮肤 细胞系 肿瘤 肿瘤 实验性 姜黄素 西达本胺 细胞凋亡 细胞增殖 药物协同作用 

分 类 号：R739.5[医药卫生—肿瘤]