蛇床子素对肝癌Huh7细胞增殖、凋亡及放射敏感性的影响及机制研究  

Effects of Osthole on Proliferation,Apoptosis and Radiosensitivity of Hepatocellular Carcinoma Huh7 Cells and Underlying Mechanism

作　　者：杨士杰[1] 刘春盛[1] 高丽环 Yang Shijie;Liu Chunsheng;Gao Lihuan(Department of Radiotherapy and Chemotherapy,Jizhong Energy Xingtai Mining Group General Hospital,Hebei Xingtai 054000,China)

机构地区：[1]冀中能源邢台矿业集团总医院放化疗科,河北邢台054000

出　　处：《中国药师》2020年第11期2094-2099,共6页China Pharmacist

基　　金：2011年邢台市科学技术研究与发展指导计划项目(编号:2011ZC155)。

摘　　要：目的:探讨蛇床子素对人肝癌Huh7细胞增殖、凋亡及放射敏感性的影响,并初步探究其作用机制。方法:采用不同浓度(0,2.5,5,10,20,40μg·ml^-1)的蛇床子素干预人肝癌Huh7细胞,噻唑蓝(MTT)法检测蛇床子素对肝癌Huh7细胞增殖的影响,并计算半数抑制浓度(IC50);10μg·ml^-1蛇床子素联合不同照射剂量(0,2,4,6,8 Gy)的X射线处理Huh7细胞,MTT检测细胞增殖能力,筛选照射剂量;克隆形成实验检测Huh7细胞放射敏感性变化,分别采用10μg·ml^-1蛇床子素、4 GyX射线照射及两者联合干预Huh7细胞,流式细胞术检测Huh7细胞凋亡率,Western Blot检测Huh7细胞中剪切的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved caspase-3)和剪切的含半胱氨酸的天冬氨酸蛋白水解酶9(cleaved caspase-9)的表达水平。结果:蛇床子素对肝癌Huh7细胞增殖有明显抑制作用,IC50为20.14μg·ml^-1;与单纯照射组相比,蛇床子素联合不同剂量X射线照射下均能够明显抑制细胞存活率,筛选出4 Gy剂量用于后续放射实验。与单纯照射组相比,蛇床子素联合照射组Huh7细胞的放射敏感性、Huh7细胞凋亡率、cleaved caspase-3和cleaved caspase-9蛋白表达显著升高(P<0.05)。结论:蛇床子素能够抑制肝癌Huh7细胞增殖,促进细胞凋亡,并增强细胞对放射的敏感性,其作用机制可能与蛇床子素上调cleaved caspase-3和cleaved caspase-9蛋白表达有关。Objective:To investigate the effects of osthole on the proliferation,apoptosis and radiosensitivity of human hepatoma Huh7 cells,and to explore its mechanism.Methods:Different concentrations (0,2.5,5,10,20,40μg·ml^-1) of osthole were used to intervene human hepatoma Huh7 cells.The effect of osthole on the proliferation of hepatoma Huh7 cells was detected by MTT assay,and the half inhibitory concentration (IC50) was calculated.10μg·ml^-1 Osthole combined with different radiation doses (0,2,4,6,8 Gy) was used to treat Huh7 cells,MTT was used to detect the cell proliferation ability,and the irradiation dose was screened.The colony formation assay was used to detect the radiosensitivity of Huh7 cells respectively treated with 10μg·ml^-1osthole,4 Gy irradiation and the combination of two,and cleaved caspase-3 and cleaved caspase-9 in Huh7 cells were detected by Western blot.Results:Osthole significantly inhibited the proliferation of hepatoma Huh7 cells with IC50of 20.14μg·ml^-1.Compared with the irradiated group,osthole combined with different doses of X-rays could significantly inhibit the cell viability.The dose of 4 Gy was screened out for the subsequent radioactivity experiments.The radiosensitivity of Humus cells,the apoptosis rate of Huh7 cells,and the expressions of cleaved caspase-3 and cleaved caspase-9 in osthole combined with irradiation group were significantly higher than those in the irradiation group (P<0.05).Conclusion:Osthol can inhibit the proliferation of hepatoma Huh7 cells,promote the cell apoptosis,and enhance the sensitivity of cells to radiation.The mechanism may be related to the up-regulation of cleaved caspase-3 and cleaved caspase-9 protein expressions by osthole.

关 键 词：蛇床子素 肝癌Huh7细胞 增殖 凋亡 放射敏感性 

分 类 号：R965.1[医药卫生—药理学]