MS-275联合顺铂对食管鳞状细胞癌EC9706细胞氧化损伤和凋亡的影响  

Effects of MS-275 combined with DDP on oxidative damage and apoptosis in esophageal squamous cell carcinoma EC9706 cells

作　　者：刘腾飞 王亚苹 李亚 张振坤 李喆 周馨魁 张璐玉 王文杰 刘红涛 张彦婷 关方霞 马珊珊 LIU Tengfei;WANG Yaping;LI Ya;ZHANG Zhenkun;LI Zhe;ZHOU Xinkui;ZHANG Luyu;WANG Wenjie;LIU Hongtao;ZHANG Yanting;GUAN Fangxia;MA Shanshan(College of Life Sciences,Zhengzhou University,Zhengzhou 450001)

机构地区：[1]郑州大学生命科学学院,郑州450001

出　　处：《郑州大学学报（医学版）》2019年第3期358-362,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省自然科学基金项目(182300410377);河南省高等学校重点科研项目计划(17A180016)

摘　　要：目的:探讨MS-275联合顺铂(DDP)对食管鳞状细胞癌(鳞癌) EC9706细胞氧化损伤和细胞凋亡的影响。方法:采用CCK-8法检测0. 5、1. 0、2. 0、4. 0和8. 0μmol/L MS-275和0. 25、0. 50、1. 00、2. 00和4. 00μmol/L DDP作用24、48、72 h对EC9706细胞存活率的影响。EC9706细胞分为空白对照组(不作处理)、MS-275组、DDP组和联合(MS-275+DDP)组,MS-275和DDP分别采用2. 0和1. 00μmol/L的浓度作用48 h,CCK-8法检测细胞的存活率,流式细胞仪检测细胞凋亡和细胞内活性氧(ROS)水平,相应试剂盒检测丙二醛(MDA)含量、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)活力,JC-1染色检测线粒体膜电位,Western blot检测Bax、Bcl-2和Trx蛋白的表达。结果:2. 0μmol/L MS-275和1. 00μmol/L DDP均能降低EC9706细胞存活率,升高凋亡率(P均<0. 001),增加细胞内ROS水平和MDA含量(P均<0. 001),降低线粒体膜电位以及SOD、GSH-PX活力(P均<0. 001),同时降低Bcl-2和Trx蛋白表达,增加Bax蛋白表达(P均<0. 001);两药联合具有一定的协同效应(P均<0. 001)。结论:MS-275联合DDP可诱导EC9706细胞的氧化损伤和凋亡。Aim:To investigate the effects of MS-275 combined with cis-diammine-dichloroplatinum(DDP)on oxidative damage and apoptosis in esophageal squamous cell carcinoma EC9706 cells.Methods:The effects of 0.5,1.0,2.0,4.0 and 8.0μmol/L MS-275 and 0.25,0.50,1.00,2.00 and 4.00μmol/L DDP for 24,48,72 hours on the survival rate of EC9706 cells were detected by CCK-8 assay.EC9706 cells were divided into blank control group,MS-275 group,DDP group and MS-275+DDP combination group.The survival rate of each group was detected by CCK-8 assay.Apoptosis and intracellular reactive oxygen species(ROS)levels were detected by flow cytometry;malondialdehyde(MDA)content,superoxide dismutase(SOD)activity and glutathione peroxidase(GSH-PX)activity in EC9706 cells were detected;mitochondrial membrane potential changes were detected by JC-1 staining;the expressions of Bax,Bcl-2 and Trx proteins were detected by Western blot.Results:Both MS-275 and DDP had a time-and dose-dependent inhibitory effects on the growth of EC9706 cells(P<0.001);2.0μmol/L MS-275 and 1.00μmol/L DDP could decrease cell viability and increase apoptosis rate,increase ROS level and MDA content,decrease SOD activity,GSH-PX activity and mitochondrial membrane potential.In addition,they could also inhibit Trx and Bcl-2 protein expressions and decrease the ratio of Bcl-2 to Bax in EC9706 cells(P<0.001).Furthermore,the combination of MS-275 and DDP had a synergistic effect(P<0.001).Conclusion:MS-275 combined with DDP could induce oxidative damage and apoptosis in EC9706 cells.

关 键 词：MS-275 顺铂 食管鳞状细胞癌 氧化损伤 凋亡 

分 类 号：R735.1[医药卫生—肿瘤]