鱼藤素对小鼠神经母细胞瘤细胞恶性行为的作用及其机制  

Effects of deguelin on malignant behavior of mouse neuroblastoma cells and its mechanism

作　　者：冯珏利 王瑛 常新捷 步星耀[2] FENG Jueli;WANG Ying;CHANG Xinjie;BU Xingyao(Department of Pediatric Surgery,the First Affiliated Hospital of He'nan University of Science and Technology,Luoyang 471000,China;Department of Neurosurgery,Henan People's Hospital,Zhengzhou 450000,China)

机构地区：[1]河南科技大学第一附属医院小儿外科,洛阳471000 [2]河南省人民医院神经外科,郑州450000

出　　处：《西北药学杂志》2022年第3期107-111,共5页Northwest Pharmaceutical Journal

基　　金：2017年河南省科技研发专项项目(编号:152102310136)。

摘　　要：目的探讨鱼藤素对小鼠神经母细胞瘤细胞恶性生物学行为的作用及机制。方法取处于对数生长期的Neuro-2A细胞,用不同浓度(0、0.01、0.1、1、10、100、1000 nmol·L^(-1))的鱼藤素作用48 h,用MTT法检测细胞存活率,选择鱼藤素对Neuro-2A细胞的半数抑制浓度(IC_(50))进行后续实验。将细胞随机分为对照组、鱼藤素组、LPS组和鱼藤素+LPS组,用MTT法检测细胞的相对活力,用流式细胞术检测细胞凋亡率,用RT-PCR检测Toll样受体4(toll-like receptor 4,TLR4)、核因子-κB(nuclear factor-κB,NF-κB)mRNA的表达水平,用Western blot检测细胞增殖核抗原67(antigen identified by monoclonal antibody 67,Ki67)、活化的含半胱氨酸的天冬氨酸蛋白水解酶(cleaved-caspase3)和TLR4/NF-κB通路相关蛋白的表达水平。结果用不同浓度的鱼藤素处理Neuro-2A细胞后,细胞存活率呈浓度依赖性降低。与对照组比较,鱼藤素组细胞的相对活力降低,凋亡率升高,TLR4 mRNA的相对表达量降低,cleaved-caspase3蛋白的相对表达量升高,Ki67、TLR4和p-p65蛋白的相对表达量降低(P<0.05),LPS组细胞的相对活力提高,凋亡率降低,TLR4 mRNA的相对表达量升高,cleaved-caspase3蛋白的相对表达量降低,Ki67、TLR4和p-p65蛋白的相对表达量升高(P<0.05);与鱼藤素组比较,鱼藤素+LPS组细胞的相对活力升高,凋亡率降低,TLR4 mRNA的相对表达量升高,cleaved-caspase3蛋白的相对表达量降低,Ki67、TLR4和p-p65蛋白的相对表达量升高(P<0.05);与LPS组比较,鱼藤素+LPS组细胞的相对活力降低,凋亡率升高,TLR4 mRNA的相对表达量降低,cleaved-caspase3蛋白的相对表达量升高,Ki67、TLR4和p-p65蛋白的相对表达量降低(P<0.05)。结论鱼藤素可以抑制小鼠神经母细胞瘤细胞增殖,诱导细胞凋亡,其作用可能通过抑制TLR4/NF-κB通路,调控细胞增殖、凋亡相关蛋白的表达实现。Objective To investigate the effect and mechanism of deguelin on malignant biological behavior of mouse neuroblastoma cells.Methods Neuro-2 A cells in logarithmic phase were treated with different concentrations of deguelin(0,0.01,0.1,1,10,100 and 1 000 nmol·L^(-1)) for 48 h.MTT assay was used to detect cell viability.IC_(50) of deguelin on Neuro-2 A was selected for subsequent experiments.The cells were randomly divided into control group, deguelin group, lipopolysaccharide(LPS) group and deguelin+LPS group.The relative viability of cells was measured by MTT method.The apoptosis rate of cells was detected by flow cytometry.The mRNA expression levels of Toll-like receptor 4(TLR4) and nuclear factor-κB(NF-κB) were detected by RT-PCR.The expression levels of antigen identified by monoclonal antibody 67(Ki67),cleaved-caspase3 and TLR4/NF-κB pathway associated proteins were detected by Western blot.Results After treated with different concentrations of deguelin, the cell Neuro-2 A viability decreased in a concentration-dependent manner.Compared with the control group, the relative activity of cells in the deguelin group decreased, the apoptosis rate increased, the relative expression of TLR4 mRNA decreased, the relative expression of cleaved-caspase3 protein increased, and the relative expression of Ki67,TLR4 and p-p65 proteins decreased(P<0.05).The relative activity of cells in the LPS group increased, the apoptosis rate decreased, the relative expression of TLR4 mRNA increased, the relative expression of cleaved-caspase3 protein decreased, and the relative expression of Ki67,TLR4 and p-p65 proteins increased(P<0.05).Compared with the deguelin group, the relative activity of cells in the deguelin+LPS group increased, the apoptosis rate decreased, the relative expression of TLR4 mRNA increased, the relative expression of cleaved-caspase3 protein decreased, and the relative expression of Ki67,TLR4 and p-p65 proteins increased(P<0.05).Compared with the LPS group, the relative activity of cells in the deguelin+LPS g

关 键 词：鱼藤素 神经母细胞瘤 增殖 凋亡 TLR4/NF-κB通路 

分 类 号：R965[医药卫生—药理学]