华蟾素联合多柔比星通过Fas/线粒体通路促进肺癌A549细胞凋亡  

Effect of cinobufotalin combined with doxorubicin on A549 cells apoptosis through Fas/mitochondrial pathway in treatment of lung cancer patients

作　　者：董伯升[1] 徐克友[1] 左彩莹[1] 张镇 DONG Bo-sheng;XU Ke-you;ZUO Cai-ying;ZHANG Zhen.(Department of Oncology, Zhoukou Central Hospital, Zhoukou, Henan 466000, China)

机构地区：[1]周口市中心医院肿瘤内科,河南周口466000 [2]河南省肿瘤医院麻醉科,河南郑州450008

出　　处：《临床肺科杂志》2019年第1期121-125,143,共6页Journal of Clinical Pulmonary Medicine

摘　　要：目的探究华蟾素联合多柔比星对肺癌A549细胞凋亡及Fas/线粒体通路的影响。方法用不同浓度华蟾素(0、0. 1、0. 2、0. 4、0. 8μg/m L)、不同浓度多柔比星(0、0. 25、0. 5、1. 0、2. 0μg/m L)或联合用药(多柔比星0. 5μg/m L分别与华蟾素0、0. 1、0. 2、0. 4、0. 8μg/m L联合使用)处理A549细胞。四甲基偶氮唑(MTT)法检测华蟾素联合多柔比星对A549细胞增殖的影响;流式细胞仪检测华蟾素联合多柔比星对A549细胞凋亡的影响;蛋白质印迹(Western blot)技术检测A549细胞Fas/线粒体凋亡通路相关蛋白的表达变化。结果 MTT结果显示,华蟾素和多柔比星对A549细胞均具有增殖抑制作用(P <0. 05),且呈时间-剂量依赖关系。0. 5μg/m L多柔比星分别与0. 1、0. 2、0. 4、0. 8μg/m L华蟾素联合处理下,A549细胞的增殖抑制率呈时间-剂量依赖性。(2)流式细胞仪结果显示,与空白对照组比较,单用0. 4μg/m L华蟾素或0. 5μg/m L多柔比星处理后A549细胞凋亡率显著上升(P <0. 05);与单独用药比较,两药联合处理后A549细胞凋亡率显著上升(P <0. 05)。(3)Western blot结果显示,与空白对照组比较,单用0. 4μg/m L华蟾素或0. 5μg/m L多柔比星处理后A549细胞Fas、Fas L、TNF-R1、Bax及Cyt-c表达显著上调、Bcl-2表达显著下调、裂解型Caspase-3/8/9显著增多(P <0. 05);与单独用药比较,两药联合处理后A549细胞Fas、Fas L、TNF-R1、Bax及Cyt-c表达显著上调、Bcl-2表达显著下调、裂解型Caspase-3/8/9显著增多(P <0. 05)。结论华蟾素联合多柔比星可显著促进肺癌A549细胞凋亡,其机制可能与Fas/线粒体凋亡通路强化有关。Objective To investigate the effect of cinobufotalin combined with doxorubicin on A549 cells apoptosis through Fas/mitochondrial pathway in treatment of lung cancer patients.Methods A549 cells were treated with different concentrations of cinobufotalin(0,0.1,0.2,0.4,0.8μg/mL)and different concentrations of doxorubicin(0,0.25,0.5,1,2μg/mL)or combined treatment(doxorubicin 0.5μg/mL combined with cinobufotalin 0,0.1,0.2 0.4 and 0.8μg/mL respectively).The effect of cinobufotalin combined with doxorubicin on the proliferation of A549 cells was detected by four methyl azazolazole(MTT)method.The effect of cinobufotalin combined with doxorubicin on the apoptosis of A549 cells was detected by flow cytometry.The expression changes of Fas/mitochondrial apoptosis pathway related proteins in A549 cells were detected by Western blot.Results ①MTT results showed that both cinobufotalin and doxorubicin could inhibit A549 cells proliferation(P<0.05),with time-dose dependence.The proliferation inhibition rate of A549 cells was time-dose dependent under the treatment of 0.5μg/mL of doxorubicin combined with 0.1,0.2,0.4,0.8μg/mL of cinobufotalin respectively.②Flow cytometry results showed that,compared with the blank control group,the apoptosis rate of A549 cells increased significantly(P<0.05)after treated with 0.4μg/mL cinobufotalin or 0.5μg/mL dDoxorubicin.Compared with single treatment,the apoptosis rate of A549 cells increased significantly(P<0.05)after the combined treatment.③The Western blot results showed that,compared with the blank control group,the expression of Fas,FasL,TNF-R1,Bax and Cyt-c in A549 cells was significantly up-regulated after treated with 0.4μg/mL cinobufotalin or 0.5μg/mL doxorubicin.The expression of Bcl-2 was significantly down-regulated.The cleavage type Caspase-3/8/9 increased significantly(P<0.05).Compared with single treatment,the expression of Fas,FasL,TNF-R1,Bax and Cyt-c in A549 cells was significantly up-regulated after the combined treatment,and the expression of Bcl-2 was signif

关 键 词：肺癌 A549细胞 华蟾素 多柔比星 Fas信号通路 线粒体信号通路 

分 类 号：R734.2[医药卫生—肿瘤]