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目的肠道病毒71型(Enterovirus 71,EV 71)是手足口病(hand,foot and mouth disease,HFMD)主要的病原体之一。研究RNA干扰技术抑制肠道病毒71型复制。方法以RNA干扰技术(RNAi)为干预手段,抑制病毒EV 71在人横纹肌肉瘤(rhabdomyosarcoma,RD)细胞中的复制。通过Western blot、Real-time PCR和病毒滴度3种方法验证,其抑制效果体现在感染细胞内部病毒RNA,病毒蛋白质的表达水平和培养基上清中子代病毒颗粒的数量上。结果靶向病毒基因组5’UTR和VP2的siRNAs具有明显的病毒抑制效应。结论此研究证实RNAi方法具有特异性抑制病毒复制的潜能和可行性。
Objective Enterovirus 71 (EV 71) is one of the major pathogens of hand, foot and mouth disease (HFMD). RNA interference technology was studied to inhibit enterovirus 71 replication. Methods RNA interference (RNAi) was used as an intervention to inhibit the replication of virus EV 71 in human rhabdomyosarcoma (RD) cells. The results of Western blot, Real-time PCR and virus titer showed that the inhibitory effect was manifested in the expression of viral RNA and viral proteins in the infected cells and the number of progeny virus particles in the medium supernatant. Results The siRNAs targeting the 5 ’UTR and VP2 of the viral genome had a significant viral suppression effect. Conclusion This study demonstrates that RNAi has the potential and feasibility of specifically inhibiting viral replication.