双香豆素体外抗肿瘤活性筛选及相关机制初探  

Anti-tumor activity screening and research on the primary mechanism of dicumarol in vitro

作　　者：魏静 冯跃平 郑茜 王钦 张春 WEI Jing;FENG Yue-ping;ZHENG Xi;WANG Qin;ZHANG Chun(Department of Pharmacy,Southwest Medical University,Luzhou 646000,China;Sichuan Vacotional College of Health and Rehabilitation,Zigong 643000,China)

机构地区：[1]西南医科大学药学院,四川泸州646000 [2]四川卫生康复职业学院,四川自贡643000

出　　处：《药学学报》2020年第12期2904-2910,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81001700);四川省青年科技创新研究团队研究项目(2019JDTD0016);泸州市-西南医科大学联合项目(2019LZXNYDJ49).

摘　　要：为初步探讨双香豆素抗肿瘤活性及相关机制,采用CCK-8法检测双香豆素对几种肿瘤细胞株生长的影响。以HepG2为研究对象,通过对几种生理生化指标的检测初步探讨其抗肿瘤的相关机制。结果显示,双香豆素对HepG2、Hccc-9810及MDA-MB-231细胞株的生长均呈剂量和时间依赖性的抑制作用,其中HepG2对双香豆素的敏感性最高(IC50=3.19±0.68μmol·L-1)。双香豆素可使HepG2细胞周期阻滞在S期,使Bcl-2蛋白的表达下调,cleaved caspase-9和Bax蛋白的表达升高。双香豆素可使HepG2细胞内还原型谷胱甘肽(glutathione,GSH)和超氧化物歧化酶(superoxide dismutase,SOD)含量明显降低,丙二醛(malonaldehyde,MDA)和活性氧(reactive oxygen species,ROS)水平明显升高。在低氧诱导下,双香豆素能使HepG2细胞中NAD(P)H醌氧化还原酶1[NAD(P)H quinone oxidoreductase 1,NQO1]、3-磷酸肌醇依赖性蛋白激酶1(3-phosphoinositide-dependent protein kinase 1,PDK1)及缺氧诱导因子1α(hypoxia inducible factor-1α,HIF-1α)表达量下调。上述结果表明,双香豆素可有效抑制HepG2细胞增殖,促进其周期阻滞及细胞凋亡;双香豆素可能通过抑制HepG2细胞中PDK1和NQO1的表达,导致HIF-1α下调、ROS聚集,从而产生细胞氧化应激促使HepG2细胞凋亡。To study the anti-tumor activities and the related mechanisms of dicumarol,the CCK-8 method was used to identify anti-tumor activities of dicumarol.HepG2 cells were used to explore the anti-tumor mechanisms by measuring several physiological and biochemical indexes.The results show that dicumarol can significantly inhibit the growth of HepG2,Hccc-9810 and MDA-MB-231 cell lines in a dose-dependent and time-dependent manner,with HepG2 cells showing the greatest sensitivity to dicumarol(with an IC50 value of 3.19±0.68μmol·L-1 at 48 h).Dicumarol arrested the cell cycle at S phase and down-regulated the expression of anti-apoptotic protein Bcl-2 while promoting the expression of the pro-apoptotic proteins Bax and cleaved caspase-9.Dicumarol significantly decreased the levels of glutathione(GSH)and superoxide dismutase(SOD)in HepG2 cells,and increased the levels of malonaldehyde(MDA)and reactive oxygen species(ROS).Dicumarol also down-regulated the protein levels of NAD(P)H quinone oxidoreductase 1,3-phosphoinositide-dependent protein kinase 1,and hypoxia inducible factor-1αunder hypoxic conditions.The above results show that dicumarol can inhibit the proliferation of HepG2 cells and induce cycle arrest and apoptosis.Dicumarol may down-regulate the expression of HIF-1αby inhibiting the activity of NQO1 and PDK1,which leads to the accumulation of ROS,thereby generating oxidative stress and inducing apoptosis in HepG2 cells.

关 键 词：双香豆素 HEPG2 凋亡 细胞周期 氧化应激 

分 类 号：R966[医药卫生—药理学]