氧化苦参碱通过下调程序性细胞死亡1蛋白表达逆转顺铂耐药非小细胞肺癌细胞A549/Cis的顺铂耐药性  

Oxymatrine reverses cisplatin(Cis) resistance in non-small-cell lung cancer A549/Cis cells by down-regulating programmed cell death 1 protein

作　　者：姬颖华[1] 杨晓煜[2] 孟祥丽[1] 王瑾 路平[1] JI Ying-hua;YANG Xiao-yu;MENG Xiang-li;WANG Jin;LU Ping(Department of Oncology,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,China;Department of Pathology,School of Basic Medicine,Xinxiang Medical College,Xinxiang 453000,China)

机构地区：[1]新乡医学院第一附属医院肿瘤科,河南卫辉453100 [2]新乡医学院基础医学院病理学系,河南新乡453003

出　　处：《中国药理学与毒理学杂志》2020年第10期743-749,共7页Chinese Journal of Pharmacology and Toxicology

基　　金：河南省医学科技攻关计划项目(2018020369)。

摘　　要：目的研究氧化苦参碱(Oxy)对顺铂(Cis)耐药的非小细胞肺癌细胞A549(A549/Cis细胞)耐药性影响和机制。方法 A549或A549/Cis细胞单独用Cis 2,4,8,16,32和64 mg·L^-1或与Oxy 20 mg·L^-1联合处理24 h,MTT法检测细胞存活抑制率和耐药指数(RI)。A549/Cis细胞分为细胞对照、Cis 5 mg·L^-1、Oxy 20 mg·L^-1、Cis 5 mg·L^-1+Oxy 20 mg·L^-1、Cis 5 mg·L^-1+空载对照质粒(pc-NC)、Cis 5 mg·L^-1+程序性细胞死亡蛋白1(PD1)过表达质粒(pc-PD1)和Cis 5 mg·L^-1+Oxy 20 mg·L^-1+pc-PD1组。流式细胞术检测A549/Cis细胞凋亡,RT-qPCR检测A549/Cis细胞中PD1 m RNA表达,Western印迹法检测A549/Cis细胞中PD1蛋白表达水平。结果 Cis对A549细胞的细胞半数抑制浓度(IC50)为10.25 mg·L^-1,对A549/Cis细胞的IC50为36.33 mg·L^-1,RI=3.54。Cis+Oxy 20 mg·L^-1组A549/Cis细胞IC50为10.67 mg·L^-1,RI为1.04。与A549/Cis细胞对照组相比,Cis 5 mg·L^-1组细胞凋亡率无明显变化,Oxy 20 mg·L^-1组细胞凋亡率显著增加(P<0.05);与Cis 5 mg·L^-1组相比,Cis 5 mg·L^-1+Oxy 20 mg·L^-1组细胞凋亡率显著增加(P<0.01),Cis5 mg·L^-1+pc-PD1组细胞凋亡率显著减少(P<0.01);与Cis 5 mg·L^-1+Oxy 20 mg·L^-1组相比较,Cis 5 mg·L^-1+Oxy 20 mg·L^-1+pc-PD1组细胞凋亡率显著减少(P<0.01)。与A549/Cis细胞对照组相比,Cis 5 mg·L^-1组A549/Cis细胞中PD1 mRNA表达无明显变化,Oxy 20 mg·L^-1组A549/Cis细胞中PD1 mRNA表达显著下调(P<0.05);与Cis 5 mg·L^-1组相比,Cis 5 mg·L^-1+Oxy 20 mg·L^-1组A549/Cis细胞中PD1 mRNA表达显著下调(P<0.01),Cis 5 mg·L^-1+pc-PD1组A549/Cis细胞中PD1 mRNA表达显著上调(P<0.01);与Cis 5 mg·L^-1+Oxy 20 mg·L^-1组相比较,Cis 5 mg·L^-1+Oxy 20 mg·L^-1+pc-PD1组A549/Cis细胞中PD1mRNA表达显著上调(P<0.01)。与A549/Cis细胞对照组相比,Cis 5 mg·L^-1组A549/Cis细胞中PD1蛋白表达无明显变化,Oxy 20 mg·L^-1组A549/Cis细胞中PD1蛋白表达显著下调(P<0.05);与Cis 5 mg·L^-1组相比,Cis 5 mg·L^-1+pc-NC组A549/Cis细胞中PD1蛋白OBJECTIVE To investigate the effect and mechanism of oxymatrine(Oxy)on cisplatin(Cis)resistant non-small-cell lung cancer A549(A549/Cis)cells.METHODS A549 or A549/Cis cells were treated with Cis 2,4,8,16,32 and 64 mg·L^-1 alone or in combination with Oxy 20 mg·L^-1 for 24 h.Cell survival inhibitory rate and drug resistance index(RI)were measured by MTT assay.A549/Cis cells were divided into cell control group,Cis 5 mg·L^-1 group,Oxy 20 mg·L^-1 group,Cis 5 mg·L^-1+Oxy20 mg·L^-1 group,Cis mg·L^-1+no-load control plasmid(pc-NC)group,Cis 5 mg·L^-1+programmed cell death 1 protein(PD1)overexpressed plasmid(pc-PD1)group,and Cis 5 mg·L^-1+Oxy 20 mg·L^-1+pc-PD1 group.Apoptosis of A549/Cis cells was detected by flow cytometry,PD1 mRNA expression of A549/Cis cells was detected by RT-qPCR,and the relative PD1 protein expression level in A549/Cis cells was detected by Western blotting.RESULTS The inhibitory concentration 50(IC50)of Cis was10.25 mg·L^-1 for A549 cells and 36.33 mg·L^-1 for A549/Cis cells(RI was 3.54).In Oxy 20 mg·L^-1 group,IC50 of Cis on A549/Cis cells was 10.67 mg·L^-1(RI was 1.04).Compared with cell control group,the apoptosis rate in Cis 5 mg·L^-1 group did not change significantly,but was significantly increased Oxy20 mg·L^-1 group(P<0.05).Compared with Cis 5 mg·L^-1 in group,the apoptosis rate was significantly increased in Cis 5 mg·L^-1+Oxy 20 mg·L^-1 group(P<0.01),but was significantly decreased in Cis 5 mg·L^-1+pc-PD1 group(P<0.01).Compared with Cis 5 mg·L^-1+Oxy 20 mg·L^-1 group,Cis 5 mg·L^-1+Oxy 20 mg·L^-1+pc-PD1 group had a significantly decreased apoptosis rate(P<0.01).Compared with cell control group,there was no significant change in PD1 mRNA expression in A549/Cis cells in Cis5 mg·L^-1 group,but PD1 mRNA expression was significantly down-regulated in A549/Cis cells in Oxy20 mg·L^-1 group(P<0.05).Compared with Cis 5 mg·L^-1 group,PD1 m RNA expression was significantly down-regulated in A549/Cis cells in Cis 5 mg·L^-1+Oxy 20 mg·L^-1 group(P<0.01),but significantly up-

关 键 词：氧化苦参碱 程序性细胞死亡蛋白1 非小细胞肺癌 细胞增殖 细胞凋亡 

分 类 号：R285[医药卫生—中药学]