双氢青蒿素通过PI3K/Akt通路逆转肺癌顺铂耐药A549/DDP细胞的分子机制  

Molecular Mechanism of Dihydroartemisinin Reversing Cisplatin Resistance of Human Lung Cancer Cell Line A549/DDP through PI3K/Akt Pathway

作　　者：孙卉 陈晓静 刘丽 张真 Sun Hui;Chen Xiaojing;Liu Li;Zhang Zhen(The First Department of Tumor Radiotherapy,Qingdao Central Hospital,Shandong Qingdao 266042,China;Department of Stereotactic Radiotherapy,Qingdao Central Hospital,Shandong Qingdao 266042,China)

机构地区：[1]青岛中心医院肿瘤放射治疗一科,山东青岛266042 [2]青岛中心医院肿瘤立体定向放疗科,山东青岛266042

出　　处：《中国药师》2021年第6期1013-1017,共5页China Pharmacist

摘　　要：目的:研究双氢青蒿素通过磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶(PI3K/Akt)通路逆转肺癌顺铂耐药A549/DDP细胞的分子机制。方法:体外培养人肺癌耐顺铂株(A549/DDP),给予不同浓度双氢青蒿素(20,40,80,160,200,250,300μmol·L^(-1))处理48 h后,通过MTT法检测双氢青蒿素对A549/DDP细胞增殖影响,选取双氢青蒿素最佳实验浓度及在不同浓度DDP(0,20,40,60,80,100μmol·L^(-1))作用下观察并计算顺铂对A549/DDP细胞IC50和双氢青蒿素对A549/DDP逆转倍数;通过流式细胞术检测各组细胞凋亡率,Western blot法检测各组细胞中凋亡相关因子B淋巴细胞瘤-2基因(Bcl-2)和半胱氨酸蛋白酶3(Caspase3)及PI3K/Akt通路相关蛋白PI3K、AKT、磷酸化蛋白激酶B(p-Akt)的表达。结果:随双氢青蒿素浓度升高,细胞增殖抑制率依次升高(P<0.05),且具有浓度依赖性,IC10(32.07±1.04)μmol·L^(-1)是双氢青蒿素为最佳逆转耐药浓度;随DDP浓度的升高,双氢青蒿素A549/DDP细胞增殖抑制率随之升高(P<0.05),DDP+双氢青蒿素组顺铂对A549/DDP的IC50为(26.42±1.23)μmol·L^(-1),顺铂组为(58.16±1.32)μmol·L^(-1),双氢青蒿素的逆转耐药倍数为2.21;与对照组相比,DDP组、双氢青蒿素组、DDP+双氢青蒿素组细胞凋亡率、caspase-3表达显著升高(P<0.05),Bcl-2、PI3K、p-Akt/Akt表达显著降低(P<0.05);与DDP组、双氢青蒿素组相比,DDP+双氢青蒿素组细胞凋亡率、caspase-3表达显著升高(P<0.05),Bcl-2、PI3K、p-Akt/Akt表达显著降低(P<0.05)。结论:双氢青蒿素通过抑制PI3K/Akt通路促进A549/DDP细胞凋亡,在一定程度上可逆转肺癌A549/DDP细胞株对顺铂的耐药性。Objective:To study the molecular mechanism of dihydroartemisinin reversing cisplatin resistance of human lung cancer cell line A549/DDP through phosphatidyl-inositol-3’-kinase/protein kinase B(PI3K/Akt)pathway.Methods:Human lung cancer cell line(A549/DDP)was cultured in vitro.After 48-hour treatment with different concentrations of dihydroartemisinin(20,40,80,160,200,250 and 300μmol·L^(-1)),the effect of dihydroartemisinin on the proliferation of A549/DDP cells was detected by MTT method,the best experimental concentration of dihydroartemisinin and the reversal ratios of cisplatin to A549/DDP cell IC50and dihydroartemisinin to A549/DDP were observed and calculated at different concentrations of DDP(0,20,40,60,80 and 100μmol·L^(-1));the apoptosis rate of each group was detected by flow cytometry,the expressions of apoptosis related factor B-cell lymphoma-2(Bcl-2)and caspase-3 and PI3K/Akt pathway related proteins PI3K,Akt and phosphorylated protein kinase B(p-Akt)of each group were detected by Western blot.Results:With the increase of dihydroartemisinin concentration,the inhibition rate of cell proliferation increased in turn(P<0.05),suggesting a concentration-dependent manner,and IC10(32.07±1.04)μmol·L^(-1)was the best concentration of dihydroartemisinin to reverse drug resistance;with the increase of DDP concentration,the inhibition rate of dihydroartemisinin on A549/DDP cell proliferation increased(P<0.05),the IC50of cisplatin to A549/DDP in DDP+dihydroartemisinin group was(26.42±1.23)μmol·L^(-1),and in cisplatin group,it was(58.16±1.32)μmol·L^(-1),and the reverse resistance of dihydroartemisinin was 2.21;compared with those in the control group,the apoptosis rate and caspase-3 expression of DDP group,dihydroartemisinin group and DDP+dihydroartemisinin group increased significantly(P<0.05),and the expressions of Bcl-2,PI3K and p-Akt/Akt were significantly decreased(P<0.05);compared with those in DDP group and dihydroartemisinin group,the apoptosis rate and caspase-3 expression of DDP+dihydroartem

关 键 词：肺癌 顺铂耐药 双氢青蒿素 磷脂酰肌醇激酶/蛋白激酶B通路 

分 类 号：R965.1[医药卫生—药理学]