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
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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. 0 μmol/L MS-275 and 0. 25,0. 50,1. 00,2. 00 and 4. 00 μmol/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. 0 μmol/L MS-275 and 1. 00 μmol/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.
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. 0 μmol/L MS-275 and 0. 25,0. 50,1. 00,2. 00 and 4. 00 μmol/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. 0 μmol/L MS-275 and 1. 00 μmol/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.
引文
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[2]DASARI S,TCHOUNWOU PB.Cisplatin in cancer therapy:molecular mechanisms of action[J].Eur J Pharmacol,2014,740:364
[3]KNIPSTEIN J,GORE L.Entinostat for treatment of solid tumors and hematologic malignancies[J].Expert Opin Investig Drugs,2011,20(10):1455
[4]刘腾飞,周建康,黄团结,等.MS-275对食管鳞癌KYSE-70细胞存活、细胞周期、凋亡及迁移的影响[J].郑州大学学报(医学版),2018,53(5):547
[5]RIVERA-DEL VALLE N,CHENG TW,IRWIN ME,et al.Combinatorial effects of histone deacetylase inhibitors(HDACi),vorinostat and entinostat,and adaphostin are characterized by distinct redox alterations[J].Cancer Chemother Pharmacol,2018,81(3):483
[6]FEINGOLD PL,SURMAN DR,BROWN K,et al.Induction of thioredoxin-interacting protein by a histone deacetylase inhibitor,entinostat,is associated with dna damage and apoptosis in esophageal adenocarcinoma[J].Mol Cancer T-her,2018,17(9):2013
[7]李贞,高丽萍.氧化应激在顺铂肾毒性中的作用[J].广东医学,2010,31(19):2600
[8]ZHANG YT,ZHU XZ,HUANG TJ,et al.beta-Carotene synergistically enhances the anti-tumor effect of 5-fluorouracil on esophageal squamous cell carcinoma in vivo and in vitro[J].Toxicol Lett,2016,261:49
[9]梁丹阳,戴汉川.PINK1/Parkin通路在线粒体自噬氧化损伤中的作用[J].中国细胞生物学学报,2018,40(1):1
[10]MARTINOU JC,YOULE RJ.Mitochondria in apoptosis:Bcl-2 family members and mitochondrial dynamics[J].Dev Cell,2011,21(1):92
[11]PAN WT,YANG JY,WEI JY,et al.Functional BCL-2 regulatory genetic variants contribute to susceptibility of esophageal squamous cell carcinoma[J].Sci Rep,2015,5:2045
[12]LU J,HOLMGREN A.The thioredoxin antioxidant system[J].Free Radic Biol Med,2014,66:75
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