摘要
肠道病毒A71型(Enterovirus A71,EV-A71)是手足口病的重要病原体,为研究EV-A71感染人扁桃体上皮细胞后对细胞凋亡和细胞周期的影响,确定ERK1/2、JNK1/2、PI3K/Akt和含半胱氨酸的天冬氨酸蛋白水解酶(Cysteinyl aspartate specific proteinase,Caspase)的作用,本文以人扁桃体上皮细胞系UT-SCC-60B为细胞模型,CCK-8试剂盒检测EV-A71对UT-SCC-60B的抑制率、流式细胞仪检测EV-A71感染组和抑制剂处理组的凋亡和细胞周期、Caspase活力检测试剂盒测定Caspase-3,Caspase-8,Caspase-9活力。EV-A71以感染剂量和感染时间依赖方式抑制UT-SCC-60B增殖;EV-A71感染致UT-SCC-60B发生细胞凋亡,抑制ERK1/2、JNK1/2和PI3K/Akt能够降低UT-SCC-60B细胞凋亡比例;EV-A71感染UT-SCC-60B后发生S期阻滞,抑制ERK1/2、JNK1/2、PI3K/Akt和Caspase阻止UT-SCC-60B发生S期阻滞;EV-A71感染UT-SCC-60B能够活化Caspase-3,Caspase-8,Caspase-9且ERK1/2、JNK1/2和PI3K/Akt调控Caspase-3,Caspase-8,Caspase-9活力。因此,EV-A71能够导致人扁桃体上皮细胞UT-SCC-60B发生凋亡和S期阻滞,并且ERK1/2、JNK1/2、PI3K/Akt和Caspase参与凋亡和S期阻滞的调控。
Enterovirus-A71(EV-A71) is the major pathogen of hand, foot and mouth disease.To investigate the apoptosis and cell cycle in EV-A71-infected human tonsillar epithelial cells and determine the roles of ERK1/2,JNK1/2, PI3 K/Akt and Caspase, the human tonsillar epithelial cell line UT-SCC-60B was chosen as a model.Cell Counting kit-8(CCK-8) was used to detect the inhibition of UT-SCC-60B cells caused by EV-A71 infection. Apoptosis and cell cycles were detected using flow cytometry in EV-A71-infected groups and inhibitortreated groups. Activities of Caspase-3, Caspase-8 and Caspase-9 were measured using Caspase detection kits.Results showed that proliferation of UT-SCC-60B cells was inhibited by EV-A71 in dose-and time-dependent manners. UT-SCC-60B cells underwent apoptosis upon EV-A71 infection,and the apoptosis rate was decreased by inhibition of the ERK1/2, JNK1/2 and PI3 K/Akt pathways. EV-A71 infection induced S-phase arrest in UT-SCC-60B cells, and the percentage of the S phase also declined upon inhibition of ERK1/2, JNK1/2,PI3 K/Akt and Caspase expression. EV-A71 induced the activities of Caspase-3, Caspase-8 and Caspase-9, and these activities were regulated by the ERK1/2, JNK1/2 and PI3 K/Ak1 pathways. In conclusion, apoptosis and S-phase arrest were induced in EV-A71-infected UT-SCC-60B cells, and ERK1/2, JNK1/2, PI3 K/Akt and Caspase pathways regulated apoptosis and cell-cycle arrest.
作者
谢广成
苏萌
于莉
李丹
潘靖丹
王宏
章青
杜娈英
段招军
XIE Guangcheng;SU Meng;YU Li;LI Dan;PAN Jingdan;WANG Hong;ZHANG Qing;DU Luanying;DUAN Zhaojun(Department of Palhogenic Biology,Chengde Medical University,067000,China;Department of Infection Management,Affiliated Hospital of Chifeng University,Chifeng 024005,China;National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)
出处
《病毒学报》
CAS
CSCD
北大核心
2019年第1期77-84,共8页
Chinese Journal of Virology
基金
国家自然科学基金青年科学基金(项目号:81702008)
题目:EV71衣壳蛋白经TLR2/TLR4异源二聚体活化细胞因子反应的研究
河北省自然科学基金青年科学基金(项目号:H2018406024)
题目:TLR2募集TLR1和TLR6抗EV71感染的分子机制研究
河北省高等学校青年拔尖人才计划(项目号:BJ2017008)
题目:TLR2异源二聚体识别EV71启动天然免疫应答的研究
承德医学院高层次人才科研启动基金(项目号:201702)
题目:承德市婴幼儿群体中重要呼吸道感染病毒的流行特征研究~~