麦角甾醇与顺铂联合用药协同抑制人肺癌A549细胞增殖  

Combination of ergosterol and cisplatin for synergistic inhibition of proliferation of human lung cancer A549 cells

作　　者：吴梅佳 黄挺 米完完 应园园 王航利 张梦迪 黄绳武[1] WU Mei-jia;HUANG Ting;MI Wan-wan;YING Yuan-yuanl;WANG Hang-li;ZHANG Meng-di;HUANG Sheng-wu(College of Pharmaceutical Science,Zhejiang Chinese Medical University,Hangzhou 311402,China;Department of General Surgery,Hangzhou Red Cross Hospital,Hangzhou 310003,China)

机构地区：[1]浙江中医药大学药学院,浙江杭州311402 [2]杭州红十字会医院普外科,浙江杭州310003

出　　处：《中国药理学与毒理学杂志》2018年第6期433-440,共8页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金资助项目(81473361)~~

摘　　要：目的研究麦角甾醇(ERG)与顺铂联合用药对人肺癌A549细胞增殖的抑制作用。方法设细胞对照组、ERG 15.75~252μmol·L^(-1)组、顺铂16.75~268μmol·L^(-1)组及ERG 15.75~252μmol·L^(-1)+顺铂33.5μmol·L^(-1)组,分别作用12,24,48和72 h,MTT法检测细胞存活。另设细胞对照组,ERG 31.5μmol·L^(-1)组、顺铂33.5μmol·L^(-1)组及ERG 31.5μmol·L^(-1)+顺铂33.5μmol·L^(-1)组,作用24 h后,划痕实验检测细胞迁移能力;Hoechst33258染色观察细胞凋亡;流式细胞术测定细胞凋亡率和细胞周期;Western蛋白质印迹法检测活化胱天蛋白酶3和活化聚腺苷酸二磷酸核糖转移酶1(PARP-1)的表达。结果 ERG 15.75~252μmol·L^(-1)+顺铂33.5μmol·L^(-1)各浓度组细胞存活抑制率相较于ERG等浓度15.75~252μmol·L^(-1)及顺铂(33.5μmol·L^(-1))组均有不同程度的提高。ERG 31.5μmol·L^(-1)+顺铂33.5μmol·L^(-1)组出现凋亡样的细胞数量及凋亡率明显高于二者单用(P<0.01)。与细胞对照组相比,其余各组显著上调活化胱天蛋白酶3的表达,ERG+顺铂组活化的PARP-1蛋白片段显著增加(P<0.01)。结论 ERG与顺铂两药联用能显著抑制A549细胞增殖和迁移能力,阻滞细胞G2/M期分裂,诱导并激活PARP-1依赖的细胞凋亡。OBJECTIVE To investigate the inhibitory effect of ergosterol（ERG） combined with cisplatin on the proliferation of human lung cancer A549 cells.METHODS A549 cells were treated with ERG（15.75-252 μmol·L^-1）,cisplatin（16.75-268 μmol·L^-1） and ERG（15.75-252 μmol·L^-1）＋cisplatin（33.5 μmol·L^-1） for 12,24,48 and 72 h,respectively.MTT method was used to detect cell survival.After 24 h treatment of cel control,ERG（31.5 μmol·L^-1）,cisplatin（33.5 μmol·L^-1） and ERG（31.5 μmol·L^-1）combined with cisplatin（33.5 μmol·L^-1） groups to detect cell migration ability,Hoechst33258 apoptosis staining test was used to detect the cell apoptosis in each group.Flow cytometry was used to determine the apoptosis rate and cell cycle of A549 cells.Western blotting was used to detect the expressions of cleaved caspase 3 and cleaved poly（ADP-ribose） polymerase-1（PARP-1）.RESULTS Compared with ERG（15.75-252 μmol·L^-1） group and cisplatin（33.5 μmol·L^-1） group,the inhibitiory rate on A549 cells of ERG（15.75-252 μmol·L^-1） combined with cisplatin（33.5 μmol·L^-1） group was increased.The number of apoptosis cel s and apoptosis rate of ERG（31.5 μmol·L^-1） combined with cisplatin（33.5 μmol·L^-1）group were significantly higher than those of the two single drug treatment groups（P〈0.01）.Compared with the cell control group,Western blotting showed that ERG,cisplatin and ERG combined with cisplatin groups significantly up-regulated the expression of cleaved caspase 3.ERG combined with cisplatin group significantly increased cleaved PARP-1 protein fragment（P〈0.01）.CONCLUSION The combination of ERG and cisplatin can significantly inhibit the growth and migration of A549 cells,block G2/M cell division,and induce and activate PARP-1-dependent apoptosis.

关 键 词：麦角甾醇 顺铂 细胞凋亡 细胞周期 A549细胞 

分 类 号：R979.1[医药卫生—药品]