相关基因差异表达研究

时间：2016-03-01 14:54

        Objective To explore the different apoptotic gene expressions and apoptotic signaling transduction of human rhabdomyosarcoma (RD) cells infected by enterovirus 71 (EV71) in different stage.Methods The survival of EV71-infected RD cells was observed by trypan blue staining.The apoptotic morphology and rates of RD cells were surveyed and detected by Annexin V-FITC/PI staining and flow cytometry,respectively.PCR array was employed to analyze 88 apoptotic gene expressions from EV71-infected RD cells at 8 h and 20 h postinfection (p.i),respectively.Results After RD cells were infected with EV71 (MOI =5) at 8 h p.i,the viability was significantly decreased.Flow cytometry data demonstrated that the apoptotic rates of EV71-infected RD cells had increased to 18.0％ and 19.1％ at 20 h p.i in early and later stage respectively.RT-PCR array studies revealed significant variations in the expression of apoptotic genes.Among 88 genes,only the expression of IFN-β1 was upregulated 5.22 folds,whereas 47 genes including ACIN1,Akt,Apaf1,caspase and CIDEB were found to be downregulated that were lower than 2 folds at 8 h p.i.However,28 genes including FasL,CD40L,TNF,caspase-10 and caspase-3 were induced more than 2 folds after EV71 infection at 20 h.Conclusion The downregulation of apoptosis-related genes is associated with viral apoptosis-suppressing effect in RD cells in the early stage of EV71 infection.The death receptor signaling pathways including Fas/FasL and TNF/TNFR are activated to induce cell apoptosis in the late stage of EV71 infection.Moreover,host cell can also inhibit apoptosis by regulating signal pathway of CD40/CD40L,NF-κB/RelA and PI3K/Akt activation.

        目的 研究肠道病毒71型(EV71)感染人横纹肌肉瘤(RD)细胞后不同时期凋亡相关基因差异表达及信号通路转导机制.方法 锥虫蓝染色观察EV71感染RD细胞的存活率,Annexin V-FITC/PI双染色法检测病毒感染后凋亡细胞形态学及凋亡发生率,PCR芯片分析病毒感染RD细胞8h和20h后88个凋亡相关基因的表达情况.结果 EV71(MOI=5)感染RD细胞8h后存活率显著下降.流式细胞仪检测显示病毒感染20 h后早期凋亡及晚期凋亡细胞分别上升至18.0％和19.1％.PCR芯片检测88个凋亡相关基因,EV71感染8h后,除IFN-β1上调5.22倍外,ACIN1、Akt、Apaf1、caspase、CIDEB等47个基因发生明显下调.病毒感染20 h后,FasL、CD40L、TNF、caspase-10、caspase-3等28个基因发生2倍以上上调.结论 EV71感染早期RD细胞凋亡相关基因表达下调与病毒凋亡抑制效应相关,而感染晚期病毒可激活Fas/FasL、TNF/TNFR死亡受体信号通路诱导细胞凋亡.同时宿主细胞也通过调节CD40/CD40L、NF-κB/RelA、PI3 K/Akt等信号通路途径抑制凋亡作用.

携康国际是中国最早从事海外医疗行业的公司之一，也是为国内患者提供出国看病、国际远程会诊、国内外早癌高端筛查体检、心脏疾病治疗等全方位信息咨询与服务的权威机构。八年来，携康国际已经帮助上千名患者实现“健康之旅”，合作先进的国外医疗资源：德国慕尼黑质子中心；日本国立癌症中心、德洲会医院集团、福冈山王医院、藤田保健卫生大学医院；美国麻省总医院、丹娜法伯癌症研究院、西达赛奈医疗中心；法国居里研究所等，并且提供快捷的国外专家预约、医院预约、辅助办理出国手续、医学资料收集和翻译以及海外治疗陪同翻译等所需的全程服务。

携康国际将以更专业的态度为中国患者带来贴心服务。
出国诊疗和体检官方电话：400-067-0509。