葡萄糖剥夺增强NQO1底物β-拉帕醌的抗肿瘤作用研究  

Enhancement of antitumor effect of NQO1 substrate β-lapachone by glucose deprivation

作　　者：程学芳[1] 朱冠华[1] 熊学惠 胡娟妮 CHENG Xue-fang;ZHU Guan-hua;XIONG Xue-hui;HU Juan-ni(Department of Clinical Pharmacy,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200080)

机构地区：[1]上海交通大学附属第一人民医院临床药学科,上海200080

出　　处：《中南药学》2020年第11期1791-1794,共4页Central South Pharmacy

基　　金：国家自然科学基金青年科学基金项目(No.81503138);上海市卫生健康委员会卫生行业临床研究专项(No.20194Y0434)。

摘　　要：目的考察葡萄糖剥夺对β-拉帕醌抗肿瘤作用的影响。方法采用2-NBDG葡萄糖摄取实验检测β-拉帕醌对细胞葡萄糖摄取能力的影响;为了考察葡萄糖剥夺后对β-拉帕醌抗肿瘤作用机制的影响,采用双香豆素(DIC)、NAD^+、caspase抑制剂(z-VAD-fmk)预温孵后检测β-拉帕醌对细胞活力的影响,同时采用Western blot法检测β-拉帕醌对SIRT1蛋白表达的影响。结果 β-拉帕醌可以浓度依赖性地促进细胞对葡萄糖的摄取;完全剥夺培养基中的葡萄糖后,β-拉帕醌的细胞毒作用明显增加,IC50由先前的10.89 μmol·L^-1降低至1.80 μmol·L^-1,约降低了6倍。DIC和NAD^+预温孵60 min,能显著逆转葡萄糖剥夺条件下β-拉帕醌的细胞毒作用;而caspase抑制剂预温孵60 min不能逆转β-拉帕醌的细胞毒性。葡萄糖剥夺后,2.5 μmol·L^-1 β-拉帕醌明显下调细胞内SIRT1蛋白。结论葡萄糖剥夺可以显著增强β-拉帕醌的抗肿瘤作用,且不改变其NQO1依赖的抗肿瘤作用机制。Objective To determine the effect of glucose deprivation on the antitumor effect of β-lapachone.Methods The 2-NBDG glucose uptake test was used to detect the influence of β-lapachone on the glucose uptake capacity of cells.After the glucose deprivation,the effect of β-lapachone on the cell viability was detecting by the MTT method.To investigate the effect of glucose deprivation on the antitumor mechanism of β-lapachone,the cell viability was detected after pre-incubation with DIC,NAD^+,or caspase inhibitor.The expression of SIRT1 protein after β-lapachone treatment was detected by western blot.Results β-lapachone promoted the glucose uptake capacity of cells in a dose-dependent manner.After the glucose deprivation,the cytotoxicity of β-lapachone was significantly increased,and the value of IC50 was reduced from 10.89 μmol·L^-1 to 1.80 μmol·L^-1,about 6 times lower.The pre-incubation of DIC or NAD^+for 60 min both significantly reversed the cytotoxicity of β-lapachone under glucose deprivation.However,inhibiting caspase with pre-incubation of z-VAD-fmk for 60 min didnot reverse the β-lapachone cytotoxicity.β-lapachone (2.5 μmol·L^-1) significantly downregulated the expression of intracellular SIRT1 protein after the glucose deprivation.Conclusion Glucose deprivation can significantly enhance the antitumor effect of β-lapachone,without changing the NQO1-dependent antitumor mechanism of β-lapachone.

关 键 词：葡萄糖剥夺 β-拉帕醌 增敏 抗肿瘤作用 

分 类 号：R96[医药卫生—药理学]