反义寡核苷酸prop5在A549细胞内具有抗流感病毒活性

目的：研究硫代反义寡核苷酸prop5在细胞水平的抗流感活性及其作用机制。方法：cy3标记prop5用于考查人肺腺癌细胞A549对硫代反义寡核酸的摄取；利用实时荧光定量PCR检测流感病毒RNA拷贝数，Western印迹检测prop5对PDCD5蛋白表达和caspase-3蛋白剪切的抑制；利用间接免疫荧光和Western印迹检测prop5对病毒核糖核蛋白复合体（RNP）出核的影响；利用TUNEL检测prop5对流感病毒引起细胞凋亡的抑制作用。结果：流感病毒感染促进A549细胞摄取prop5；prop5下调感染病毒的A549细胞中PDCD5蛋白的表达，并能抑制流感病毒的复制；prop5抑制流感病毒引起的A549细胞的凋亡；prop5抑制病毒RNP出核。结论：prop5在细胞水平具有抗流感病毒活性，其作用机制可能同抑制RNP出核有关；本研究为进一步探讨宿主-病毒相互作用和抗流感药物开发奠定了基础。

Antisense Oligonucleotides prop5 Has Inhibitory Activity Against Influenza Virus in A549 Cells

Objective： To study the possible mechanism of action of antisense oligonucleotides prop5 against influenza virus. Methods： The cy3 labeled prop5 was used to test uptake of prop5 by A549 cells. Influenza viral RNA was detected by real-time quantitative PCR. Western blotting was applied to assess expression of PDCD5 protein and cleavage of caspase-3 protein. Inhibition of export of viral ribonucleoprotein complexes（RNP） from the nucleus to the cytoplasm by prop5 was test by indirect immunofluorescence microscopy and Western blotting. Inhibition of apoptosis by prop5 in A549 cells was observed by TUNEL assay. Results： Uptake of prop5 by A549 cells was enhanced after infection with influenza virus. The prop5 down-regulated expression protein level of PDCD5 in infected A549 cells, and inhibited apoptosis induce by influenza virus and export of RNP from nucleus to cytoplasm. Conclusion： The prop5 suppressed replication of influenza virus in A549 cells, which was associatied with retention of the viral RNP in the nucleus. This study provided a base for further studying host-virus interaction and an opportunity for the development of antiviral therapies.