T-2毒素抗人食管癌细胞EC1的活性及其机制  

Anticancer activity and mechanism of T-2 toxin in human esophageal carcinoma cell EC1

作　　者：马永成[1] 范霞霞 苏楠[2] MA Yongcheng;FAN Xiaxia;SU Nan(Department of Pharmacy of Henan Provincial People′s Hospital,Department of Pharmacy of Centeral China Fuwai Hospital,Zhengzhou 450003,China;College of Food and Biological Engineering,Henan University of Animal Husbandry and Economy,Zhengzhou 450011,China)

机构地区：[1]河南省人民医院药学部,郑州大学华中阜外医院药学部临床药理室,河南郑州450003 [2]河南牧业经济学院食品与生物工程学院,河南郑州450011

出　　处：《中国药理学与毒理学杂志》2023年第11期833-840,共8页Chinese Journal of Pharmacology and Toxicology

基　　金：河南省高等学校重点科研项目(21A330002);河南省医学科技攻关计划(2018020462)。

摘　　要：目的研究T-2毒素的抗癌活性,并探讨其作用机制。方法人食管癌细胞EC109和EC1、人胃癌细胞MGC-803、人肺癌细胞H460及人正常胃黏膜细胞GES-1,各细胞分别分为细胞对照组(二甲亚砜,终浓度<0.01%)和T-2毒素0.375~100 nmol·L^(-1)组,处理细胞24,48和72 h后,采用MTT法检测细胞存活率。EC1细胞分为细胞对照组和T-2毒素5和10 nmol·L^(-1)组,实时无标记动态细胞分析技术(RTCA)进一步分析EC1细胞增殖和迁移;EC1细胞分为细胞对照组和T-2毒素5,10和20 nmol·L^(-1)组,T-2毒素处理48 h后,流式细胞术检测细胞凋亡和细胞周期及活性氧(ROS)水平和线粒体膜电位(MMP),Western印迹法检测细胞色素c、多聚ADP核糖聚合酶(PARP)、剪切形式PARP、P21、γ-H2AX、P53、Bax和活化胱天蛋白酶3/7蛋白表达。结果T-2毒素作用EC109,EC1,MGC-803和H4604种人癌细胞72 h,IC50值分别为6.34,5.54,6.94和5.96 nmol·L^(-1),T-2毒素对正常胃黏膜细胞GES-1的IC50为16.4 nmol·L^(-1)。在0~25 nmol·L^(-1)浓度范围内,T-2毒素对EC1细胞增殖具有明显抑制作用,且呈浓度和时间依赖性(24 h,r=0.9204;48 h,r=0.9745;72 h,r=0.9772),此外T-2毒素亦可强烈抑制EC1细胞迁移;与细胞对照组相比,T-2毒素5,10和20 nmol·L^(-1)组EC1细胞凋亡率(P<0.01)、G2/M期细胞比例(P<0.01)、ROS含量(P<0.05,P<0.01)、低MMP细胞比例(P<0.01)、细胞质中细胞色素c含量(P<0.01)、剪切形式PARP含量(P<0.01)、P21(P<0.01)和γ-H2AX、P53、Bax及活化胱天蛋白酶3/7含量(P<0.05,P<0.01)均显著升高。结论T-2毒素通过诱导细胞凋亡和周期阻滞抑制癌细胞生长,其机制与激活线粒体凋亡通路和上调周期抑制因子P21表达有关。OBJECTIVE To investigate the anticancer potential of T-2 toxin and the underlying mechanism.METHODS Four cancer cell lines,human esophageal carcinoma cell EC109 and EC1,human gastric carcinoma cell MGC-803,and human lung cancer cell line H460,and normal gastric mucosa GES-1 cells were randomly divided into two groups:cell control(0.01%DMSO)and 0.375-100 nmol·L^(-1) T-2 toxin treatment groups.Cancer cells were treated with above-mentioned two groups for 24,48 or 72 h,MTT assay was used to measure the effect of T-2 toxin on the proliferation of the cell lines.EC1 cells were divided into cell control,T-2 toxin 5 and 10 nmol·L^(-1) groups,real time cell analysis(RTCA)was performed to further analyse the inhibiting effect of T-2 toxin on esophageal cancer cell EC1 proliferation and migration.EC1 cells were divided into cell control,T-2 toxin 5,10 and 20 nmol·L^(-1) groups.The apoptosis and cycle of cells,levels of reactive oxygen species(ROS),as well as mitochon-drial membrane potential(MMP)were analyzed by flow cytometry.The expressions of proteins,cyto-chrome c,poly(ADP-ribose)polymerase(PARP),cleaved PARP,P21,γ-H2AX,P53,Bax and activated caspase-3/7 were determined by Western blotting.RESULTS Four human cancer cell lines,EC109,EC1,MGC-803 and H460,were treated with T-2 toxin for 72 h,and the IC50 was 6.34,5.54,6.94 and 5.96 nmol·L^(-1),respectively.The IC50 of T-2 toxin against normal gastric mucosa GES-1 cells was 16.4 nmol·L^(-1).In addition,in the concentration range of 0-25 nmol·L^(-1),T-2 toxin could significantly inhibit the prolifera-tion and migration of EC1,suggesting dependence on the concentration and time(24 h,r=0.9204;48 h,r=0.9745;72 h,r=0.9772).Compared with the cell control group,the apoptosis rate of EC1 cells(P<0.01),G2/M phase cell ratio(P<0.01),ROS content(P<0.05,P<0.01),low MMP cells ratio(P<0.01),cyto-chrome c content(P<0.01),cleaved PARP content(P<0.01),p21(P<0.01),γ-H2AX(P<0.05,P<0.01),as well as the levels of p53,Bax and activated caspase-3/7(P<0.05,P<0.01)in T-25,10 and 20 nmol·L^

关 键 词：T-2毒素 细胞凋亡 细胞周期 

分 类 号：R963[医药卫生—微生物与生化药学] R99[医药卫生—药理学]