黄芪多糖对人肺癌顺铂耐药株A549/DDP细胞耐药性的影响及其机制  

Effect of astragalus polysaccharide on cisplatin resistance of human lung cancer A549/DDP cells and its mechanism

作　　者：柳叶 陈龙云 LIU Ye;CHEN Longyun(Department of Clinical Microbiology,School of Laboratory Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China;Department of Biochemistry,School of Basic Medical Sciences,Hubei University of Chinese Medicine,Wuhan 430065,China)

机构地区：[1]湖北中医药大学检验学院临床微生物学教研室,湖北武汉430065 [2]湖北中医药大学基础医学院生物化学教研室,湖北武汉430065

出　　处：《吉林大学学报（医学版）》2020年第6期1162-1168,I0004,共8页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金青年基金资助课题(31700152);湖北高校省级教学研究项目资助课题(2017353)。

摘　　要：目的:探讨黄芪多糖(APS)对人肺癌顺铂(DDP)耐药A549/DDP细胞耐药性的影响,并阐明其可能的作用机制。方法:将对数生长期A549/DDP细胞分为对照组(不经药物干预)、APS组(给予25、50、100、200和400 mg·L^-1 APS)、DDP组(给予2、4、8、16和32 mg·L^-1 DDP)及APS+DDP组(给予100 mg·L^-1 APS和2、4、8、16和32 mg·L^-1 DDP),药物处理A549/DDP细胞24 h,采用CCK-8法检测各组A549/DDP细胞增殖抑制率,并计算半数抑制浓度(IC50)。将A549/DDP细胞分为对照组(不经药物干预)、APS组(给予100 mg·L^-1 APS)、DDP组(给予11.46 mg·L^-1 DDP)及APS+DDP组(给予100 mg·L^-1 APS和11.46 mg·L^-1 DDP),药物处理细胞24 h,采用Transwell小室法检测各组A549/DDP细胞迁移情况,流式细胞术检测各组A549/DDP细胞凋亡率和线粒体膜电位,Western blotting法检测各组A549/DDP细胞中B淋巴细胞瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、P糖蛋白(P-gp)、磷脂酰肌醇3激酶(PI3K)、磷酸化磷脂酰肌醇3激酶(p-PI3K)、蛋白激酶B(AKT)和磷酸化蛋白激酶B(p-AKT)蛋白表达水平。结果:CCK-8法检测,与DDP组比较,APS+DDP组的A549/DDP细胞IC50降低。Transwell小室法检测,与对照组比较,APS组A549/DDP细胞迁移能力无明显变化;与DDP组比较,APS+DDP组A549/DDP细胞迁移数降低。流式细胞术检测,与对照组比较,APS组A549/DDP细胞凋亡率和线粒体膜电位差异无统计学意义(P>0.05),DDP组A549/DDP细胞凋亡率升高,线粒体膜电位降低(P<0.05);与DDP组比较,APS+DDP组A549/DDP细胞凋亡率升高,线粒体膜电位降低(P<0.05)。Western blotting法检测,与对照组比较,APS组A549/DDP细胞中Bcl-2和P-gp蛋白表达水平降低(P<0.05),p-PI3K/PI3K和p-AKT/AKT比值降低(P<0.05),Bax蛋白表达水平差异无统计学意义(P>0.05);与APS组比较,DDP组A549/DDP细胞中Bcl-2和P-gp蛋白表达水平降低(P<0.05),p-PI3K/PI3K和p-AKT/AKT比值降低P<0.05),Bax蛋白表达水平升高(P<0.05);与DDP组比较,APS+DDP组A549/DDP细胞中Bcl-2和P-gp蛋�Objective:To explore the effect of astragalus polysaccharide(APS)on the cisplatin(DDP)resistance of human lung cancer A549/DDP cells,and to clarify its possible mechanism.Methods:The A549/DDP cells at logarithmic growth phase were divided into control group(without drug intervention),APS groups(given 25,50,100,200,and 400 mg·L^-1 APS),DDP groups(given 2,4,8,16,and 32 mg·L^-1 DDP)and APS+DDP groups(given 100 mg·L^-1 APS and 2,4,8,16,32 mg·L^-1 DDP).After the cells were treated for 24 h,the inhibitory rates of proliferation of the A549/DDP cells in various groups were detected by CCK-8 method and the half inhibitory concentration(IC50)was caluculated.The A549/DDP cells were divided into control group(without drug intervention),APS group(given 100 mg·L^-1 APS),DDP group(given 11.46 mg·L^-1 DDP)and APS+DDP group(given 100 mg·L^-1 APS and 11.46 mg·L^-1 DDP),the cells in various groups were treated with drugs for 24 h.The migration of A549/DDP cells in various groups was detected by Transwell chamber assay;the apoptotic rates and mitochondrial membrane potentials of the A549/DDP cells in various groups were detected by flow cytometry,and the level expression levels of B-cell lymphoma 2(Bcl-2),Bcl-2-related X protein(Bax),P-glycoprotein(P-gp),phosphatidylinositol-3 kinase(PI3K),phosphorylated phosphatidylinositol-3 kinase(p-PI3K),protein kinase B(AKT)and phosphorylated protein kinase B(p-AKT)proteins in the A549/DDP cells in various groups were detected by Western blotting method.Results:The CCK-8 results showed that compared with DDP group,the IC50 of DDP in APS+DDP group was decreased.The Transwell chamber assay results showed that compared with control group,the migration number of A549/DDP cells in APS group was not significantly changed;compared with DDP group,the migration number of A549/DDP cells in APS+DDP group was decreased.The results of flow cytometry showed that compared with control group,the apoptotic rate and mitochondrial membrane potential of A549/DDP cells in APS group had no significant chang

关 键 词：黄芪多糖 A549/DDP细胞 耐药性 磷脂酰肌醇3激酶 蛋白激酶B 

分 类 号：R273[医药卫生—中西医结合]