肠道病毒71型感染人横纹肌肉瘤细胞不同时期凋亡相关基因差异表达研究  

Differentially expressed genes associated with apoptosis in human rhabdomyosarcoma cells infected by enterovirus 71

作　　者：李祥[1] 侯学伶[1] 彭宏君[1] 史梅[1] 姜庆波[1] 刘息平[1] 姚玉华[1] 何彩珍[1] 史伟峰[1] 

机构地区：[1]苏州大学附属第三医院检验科,常州213003

出　　处：《中华微生物学和免疫学杂志》2012年第8期718-724,共7页Chinese Journal of Microbiology and Immunology

基　　金：江苏省卫生厅“科教兴卫”医学重点人才项目资助

摘　　要：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，AnnexinV．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20h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-B1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20h后，FasL、CIMOL、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CIM0／CIMOL、NF—KB／RelA、P13K／Akt等信号通路途径抑制凋亡作用。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 EV7 l-infected RD cells was observed by trypan blue staining. The apoptotic mor- phology and rates of RD cells were surveyed and detected by Annexin V-FITC/PI staining and flow cytome- try, respectively. PCR array was employed to analyze 88 apoptotie gene expressions from EVTl-infccted 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 EVTl-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-~I was upregulated 5.22 folds, whereas 47 genes including ACIN1, Akt, Apafl, caspase and CIDEB were found to be downregulated that were lower than 2 folds at 8 h p.i. However, 28 genes including FasL, CIMOL, 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-KB/RelA and PI3K/Akt activation.

关 键 词：肠道病毒71型 RT—PCR芯片 FASL CD40L 细胞凋亡 

分 类 号：R73[医药卫生—肿瘤]