表没食子儿茶素没食子酸酯乙基化衍生物Y_(6)对耐阿霉素人肝癌细胞丝裂原活化蛋白激酶通路的影响  

Effect of epigallocatechin-3-gallate ethylated derivative Y_(6) on mitogen-activated protein kinase pathway in doxorubicin-resistant human hepatocellular carcinoma cells

作　　者：黄川 吴美怡 谢战洪 赵瑞强 温燕[3] HUANG Chuan;WU Meiyi;XIE Zhanhong;ZHAO Ruiqiang;WEN Yan(School of Pharmacy,Guangxi Medical University,Nanning 530021,Guangxi Province,China;Guangxi Colleges and Universities Key Laboratory of Biological Molecular Medicine Research,Nanning 530021,Guangxi Province,China;Department of Pharmacy,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi Province,China)

机构地区：[1]广西医科大学药学院,广西南宁530021 [2]广西高校生物分子医学研究重点实验室,广西南宁530021 [3]广西医科大学第一附属医院药学部,广西南宁530021

出　　处：《新乡医学院学报》2022年第11期1013-1018,共6页Journal of Xinxiang Medical University

摘　　要：目的探讨表没食子儿茶素没食子酸酯(EGCG)乙基化衍生物Y_(6)在逆转耐阿霉素(DOX)人肝癌细胞耐药过程中对丝裂原活化蛋白激酶(MAPK)信号通路中细胞外信号调节激酶1/2(ERK1/2)、c-Jun氨基末端激酶(JNK)和p38丝裂原活化蛋白激酶(p38)途径的影响。方法将对数生长期的耐DOX人肝癌细胞BEL-7404/DOX随机分为空白对照组、DOX组、DOX+维拉帕米(VER)组、DOX+EGCG组、DOX+低剂量Y_(6)组和DOX+高剂量Y_(6)组,空白对照组不加任何药物,DOX组加入终浓度为10μmol·L^(-1)的DOX,DOX+VER组加入终浓度为10μmol·L^(-1)的DOX和10μmol·L^(-1)的VER,DOX+EGCG组加入终浓度为10μmol·L^(-1)的DOX和10μmol·L^(-1)的EGCG,DOX+低剂量Y_(6)组加入终浓度为10μmol·L^(-1)的DOX和10μmol·L^(-1)的Y_(6),DOX+高剂量Y_(6)组加入终浓度为10μmol·L^(-1)的DOX和15μmol·L^(-1)的Y_(6),各组细胞加药完毕后,置于37℃、含体积分数5%CO_(2)的恒温培养箱中培养48 h,采用Western blot法检测各组细胞中磷酸化ERK1/2(p-ERK1/2)、ERK1/2、磷酸化JNK(p-JNK)、JNK、磷酸化p38(p-p38)、p38的蛋白相对表达量,并计算p-ERK1/2与ERK1/2蛋白相对表达量比值(p-ERK1/2/ERK1/2)、p-JNK与JNK蛋白相对表达量比值(p-JNK/JNK)、p-p38与p38蛋白相对表达量比值(p-p38/p38)。结果DOX组耐DOX人肝癌细胞中的p-ERK1/2/ERK1/2与空白对照组比较差异无统计学意义(P>0.05);DOX+VER组、DOX+EGCG组、DOX+低剂量Y_(6)组和DOX+高剂量Y_(6)组耐DOX人肝癌细胞中p-ERK1/2/ERK1/2均显著高于空白对照组和DOX组(P<0.05);DOX+VER组、DOX+EGCG组、DOX+低剂量Y_(6)组和DOX+高剂量Y_(6)组耐DOX人肝癌细胞中p-ERK1/2/ERK1/2比较差异均无统计学意义(P>0.05)。与空白对照组比较,DOX组、DOX+VER组、DOX+EGCG组、DOX+低剂量Y_(6)组和DOX+高剂量Y_(6)组耐DOX人肝癌细胞中p-JNK/JNK均显著增高(P<0.05);与DOX组比较,DOX+VER组、DOX+高剂量Y_(6)组耐DOX人肝癌细胞中p-JNK/JNK均显著增高(P<0.05);DOX+EGCG组、DOX+低�Objective To investigate the effect of ethylated derivative Y_(6) of epigallocatechin-3-gallate(EGCG)on the extracellular signal regulated kinase 1/2(ERK1/2),c-Jun N-terminal kinase(JNK)and p38 mitogen-activated protein kinase(p38)in the mitogen-activate d protein kinase(MAPK)signal pathway during reversing drug resistance of doxorubicin(DOX)-resistant human hepatocellular carcinoma cells.Methods The DOX-resistant human hepatocellular carcinoma BEL-7404/DOX cells in logarithmic growth period were randomly divided into blank control group,DOX group,DOX+verapamil(VER)group,DOX+EGCG group,DOX+low-dose of Y_(6) group and DOX+high-dose of Y_(6) group.The cells in the blank control group did not added any drugs,the cells in the DOX group were added with a final concentration of 10μmol·L^(-1) DOX,the cells in the DOX+VER group were added with a final concentration of 10μmol·L^(-1) DOX and 10μmol·L^(-1) VER,the cells in the DOX+EGCG group were added with a final concentration of 10μmol·L^(-1) DOX and 10μmol·L^(-1) EGCG,the cells in the DOX+low-dose of Y_(6) group were added with a final concentration of 10μmol·L^(-1) DOX and 10μmol·L^(-1) Y_(6),the cells in the DOX+high-dose of Y_(6) group were added with a final concentration of 10μmol·L^(-1) DOX and 15μmol·L^(-1) Y_(6).After drug administration,the cells in each group were cultured in a constant temperature incubator with a volume fraction of 5% CO_(2) at 37℃ for 48 hours.The relative expressions of phosphorylated ERK1/2(p-ERK1/2),ERK1/2,phosphorylated JNK(p-JNK),JNK,phosphorylated p38(p-p38)and p38 protein were detected by Western blot.The ratio of protein relative expression level of p-ERK1/2 and ERK1/2(p-ERK1/2/ERK1/2),ratio of protein relative expression level of p-JNK and JNK(p-JNK/JNK)and ratio of protein relative expression level of p-p38 and p38(p-p38/p38)were calculated.Results There was no significant difference in the p-ERK1/2/ERK1/2 of DOX-resistant human hepatocellular carcinoma cells between the DOX group and blank control group(P>0.05

关 键 词：表没食子儿茶素没食子酸酯 乙基化衍生物Y_(6) 丝裂原活化蛋白激酶 阿霉素耐药 

分 类 号：R735.7[医药卫生—肿瘤]