华蟾素增强氟尿嘧啶对食管癌TE-1细胞杀伤作用及机制研究  

Cinobufotalin enhancing the killing effect of fluorouracil on esophageal cancer TE-1 cells and its mechanisms

作　　者：刘博[1] 戚诚[1] 赵爽[2] 赵晓东[1] 刘学臣[3] 张立超[4] 石畅 LIU Bo;QI Cheng;ZHAO Shuang;ZHAO Xiao-dong;LIU Xue-chen;ZHANG Li-chao;SHI Chang(a.Department of Oncological Surgery,b.Digestive Department,c.Department of Microinvasive Surgery,the Second Hospital,Hebei Medical University,Shijiazhuang 050000,China;Department ofAnesthesiology,the Third Hospital,Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第二医院肿瘤外科,石家庄050000 [2]河北医科大学第三医院麻醉科,石家庄050000 [3]河北医科大学第二医院消化内科,石家庄050000 [4]河北医科大学第二医院微创外科,石家庄050000

出　　处：《实用药物与临床》2018年第9期973-977,共5页Practical Pharmacy and Clinical Remedies

基　　金：河北省中医药管理局科研计划项目(2017069);河北省医学科学研究重点课题计划(20160540)

摘　　要：目的观察华蟾素对氟尿嘧啶处理后的食管癌TE-1细胞增殖、细胞周期及凋亡的影响,并探讨其作用机制。方法 Western印迹检测不同浓度华蟾素干预后Bcl-2/Bax信号通路蛋白水平,MTT检测不同浓度及不同细胞培养时间华蟾素对氟尿嘧啶处理后TE-1细胞增殖的影响。流式细胞术检测华蟾素对氟尿嘧啶处理后TE-1细胞周期及凋亡的影响。结果随着Cinobufotalin浓度升高,Bcl-2、Survivin蛋白表达水平呈剂量依赖性下降,Cleaved PARP、Bax蛋白表达呈剂量依赖性上升; Cinobufotalin组TE-1细胞凋亡率高于Negative组(t=2. 236,P <0. 05),Cinobufotalin+5-FU组TE-1细胞凋亡率高于Negative Control组、Cinobufotalin组及5-FU组(P <0. 05);与Negative Control组对比,培养48 h后,Cinobufotalin组TE-1细胞的细胞周期停留在G1、G2期的比例减少,S期比例升高(P <0. 05);与Negative Control组、Cinobufotalin组及5-FU组比较,培养48 h后,Cinobufotalin+5-FU组TE-1细胞周期停留在G1、G2期的比例减少,S期比例升高(P <0. 05); MTT显示,Cinobufotalin干预后,TE-1细胞存活率呈现时间依赖性下降趋势,Cinobufotalin组TE-1细胞培养48、72、96 h后的存活率低于Negative Control组(P <0. 05); Cinobufotalin+5-FU组TE-1细胞培养48、72、96 h后存活率低于Negative Control组、Cinobufotalin组及5-FU组(P <0. 05);不同浓度梯度的华蟾素作用48 h后,Cinobufotalin组、Cinobufotalin+5-FU组TE-1细胞存活率呈现剂量依赖性下降。结论华蟾素通过Bcl-2/Bax通路降低食管癌细胞存活率,促进其凋亡,其对氟尿嘧啶活性具有增效作用。Objective To observe the effect of Cinobufotalin on the proliferation, cell cycle and apoptosis of TE-1 cells which were treated with fluorouracil, and explore its mechanism. Methods The protein level of Bcl-2/Bax signaling pathway after being treated with different concentrations of Cinobufotalin was detected by Western blotting. The proliferation of TE-1 cells which were treated with different concentrations of Cinobufotalin and for different cul-ture time was detected by MTT. The cell cycle and apoptosis of TE-1 cells were detected by flow cytometry. Results With concentrations of Cinobufotalin increasing, the expression of Bcl-2 and Survivin protein decreased in a dose-de-pendent manner, while the expression of Cleaved PARP and Bax protein increased in the same trend. The apoptosis rate of TE-1 cells in Cinobufotalin group was higher than that of Negative group （t=2. 236 ,P 〈0.05 ） ; the apoptosis rate in Cinobufotalin ＋5-FU group was higher than those of the other 3 groups （ P 〈0.05 ） . Compared with Negative Control group, Cinobufotalin group had lower proportion of cells arrested in G, phase and G2 phase and higher proportion in S phase （ P〈0.05） after being cultured for 48 h ; compared with Negative Control g roup, Cinobufotalin group and 5 -FU group, Cinobufotalin ＋5-FU group had lower proportion of cells arrested in G（ phase and G2 phase and higher propor-tion in S phase （ P 〈0.05） after being cultured for 48 h. MTT results indicated that the survival rate of TE-1 cells had a downward time-dependent trend after being treated with Cinobufotalin intervention. The survival rate of TE-1 cells in Cinobufotalin group was lower than that of Negative Control group after being curtured for 48 h,72 h and 96 h,and the survival rate in Cinobufotalin ＋5-FU group was lower than those of Negative Control group, Cinobufotalin group and 5- FU group respectively （ P〈0.05）. The survival rate of TE-1 cells in Cinobufotalin group and Cinobufotalin ＋5-FU grou

关 键 词：食管癌 华蟾素 凋亡 

分 类 号：R735.1[医药卫生—肿瘤]