淫羊藿苷联合LY294002对涎腺腺样囊性癌细胞增殖和凋亡的影响  

Effects of icariin combined with LY294002 on cell proliferation and apoptosis of salivary adenoid cystic carcinoma cells

作　　者：许伟 刘昕[1] 李萍[1] 王越[1] 魏丹[1] 吴家媛[2] 宋琦[1] 

机构地区：[1]遵义医学院附属口腔医院病理科,贵州省遵义市563000 [2]遵义医学院附属口腔医院牙体牙髓科,贵州省遵义市563000

出　　处：《广西医学》2018年第4期430-434,共5页Guangxi Medical Journal

基　　金：贵州省遵义医学院研究生教育创新计划项目[遵研创合CX字(2013)13号];贵州省遵义市红花岗区科学技术项目[遵红科合社字(2013)16号];贵州省研究生教育创新基地建设计划[黔教研合CXJD字(2014)005]

摘　　要：目的研究淫羊藿苷(ICA)联合LY294002对人涎腺腺样囊性癌肺高转移细胞株(ACC-M)细胞的促凋亡作用及机制。方法不同浓度ICA、LY294002作用ACC-M细胞24 h、48 h、72 h后采用噻唑兰比色法测定细胞的抑制率。筛选联合用药浓度并将细胞分为:A(阴性对照组,不加任何药物)、B[(LY294002抑制浓度(IC)_(50)),即阳性对照组]、C(ICA IC_(50))、D(ICA IC_(20)+LY294002 IC_(10))、E(ICA IC_(40)+LY294002 IC_(10))组;各组分别给予相应药物处理细胞48 h后,采用流式细胞仪检测各组细胞凋亡率,免疫印迹法检测各组细胞磷酸化蛋白激酶B(p-AKT)、核因子(NF)-κB蛋白表达水平。结果与A组相比,不同浓度ICA、LY294002对ACC-M细胞增殖均具有抑制作用。不同浓度ICA联合LY294002 IC_(10)均可抑制细胞增殖,抑制率与ICA浓度呈剂量依赖性(P<0.05)。干预48 h后,E组的细胞凋亡率高于B、C组(P<0.05),镜下可见各浓度药物组细胞均表现有典型的凋亡细胞形态学特征;E组p-AKT、NF-κB蛋白表达水平低于B、C组(P<0.05)。结论ICA联合LY294002可能是通过抑制p-AKT、NF-κB蛋白表达而促进ACC-M细胞的凋亡。Objective To investigate the pro-apoptotic effects and mechanism of icariin（ICA）combined with LY294002 for human salivary adenoid cystic carcinoma cell clone highly metastatic to the lung（ACC-M）.Methods Methyl thiazolyl tetrazolium assay was used to determinate the inhibition rates of ACC-M cells treated with different concentrations of ICA and LY294002 for 24,48 and 72 hours respectively.The experiment for determining the drug concentrations of combined medication was conducted,and the cells were divided into group A（negative control group,without any medication）,group B[positive control group,LY294002 inhibiting concentration（IC）_（50）],group C（ICA IC_（50））,group D（ICA IC_（20）plus LY294002 IC_（10））and group E（ICA IC_（40）plus LY294002 IC_（10））.Flow cytometry was used to detect the apoptosis rate of the cells in each group,treated with corresponding drugs for 48 hours,and Western blot to detect the protein expression levels of phosphoprotein kinase B（p-AKT）and nuclear factor kappa B（NF-κB）.Results Compared to group A,both ICA and LY294002with different concentrations had inhibitory effects on the proliferation of ACC-M cells.Combined use of ICA and LY294002 with different concentrations could inhibit the cell proliferation,and a dose-dependent relationship was observed between the inhibition rate and the ICA concentration（P0.05）.After 48 hours of intervention,the apoptosis rate in group E was higher than that in group B or C（P0.05）,and the cells in each drug group presented typical morphological features of apoptotic cells;the protein expression levels of p-AKT and NF-κB in group E were lower than those in group B or C（P0.05）.ConclusionICA combined with LY294002 might promote the apoptosis of ACC-M cells by inhibiting the protein expression of p-AKT and NF-κB.

关 键 词：涎腺腺样囊性癌 淫羊藿苷 LY294002 磷酸化蛋白激酶B 核因子-ΚB 凋亡 

分 类 号：R739.87[医药卫生—肿瘤]