华南农业大学学报2025，Vol.46Issue(2)：133-140,8.DOI:10.7671/j.issn.1001-411X.202404027
草地贪夜蛾Sf9细胞外泌体microRNA调控AcMNPV增殖的研究
Regulation of AcMNPV proliferation by exosomal microRNA in Spodoptera frugiperda Sf9 cells
摘要
Abstract
[Objective]To investigate the effect of Spodoptera frugiperda Sf9 cell exosomal microRNA(miRNA)on proliferation of Autographa californicamultiple nucleopolyhedrovirus(AcMNPV).[Method]Ex-osomes were purified using discontinuous sucrose mass fraction gradient ultracentrifugation.Then the purified exosome samples were observed under transmission electron microscopy,and conducted particle diameter analysis using nanoparticle tracking analysis.By sRNA hight throughput sequencing analysis,exosomally differentially expressed miRNAs were identified and biological pathways corresponding to their potential target genes were predicted.Cellular experiments such as mimic transfection and virus infection,as well as qPCR were performed to verify the differential expression of exosomal miRNAs,and detect the effect of the differentially expressed miRNA of sfr-miR-1a-3p on the proliferation of AcMNPV.[Result]Exosomes from Sf9 cells were successfully purified,and AcMNPV infection effectively stimulated exosome secretion from Sf9 cells.After 72 h of AcMNPV infection in Sf9 cells,a total of 11 differentially expressed miRNAs were identified in exosomal miRNAs comparing with the Rfam database,in which eight miRNAs'transcription levels showed consistent trends with the sequencing results.Potential target genes were mainly enriched in the cAMP signaling pathway,PI3K-Akt signaling pathway,cancer pathway,and human papilloma virus signaling pathway,suggesting that differentially expressed exosomal miRNAs might participate in insect innate immune responses.Overexpressing sfr-miR-1a-3p significantly promoted the expression level of the AcMNPV gene vp39.[Conclusion]This study conducts sRNA sequencing to screen differentially expressed miRNAs in AcMNPV-infected Sf9 cell exosomes,demonstrating that AcMNPV promotes infection by enhancing exosome secretion and affecting surrounding uninfected cells via miRNA sfr-miR-1a-3p transfer.This research provides an important theoretical support for understanding the mechanism by which insect exosomal miRNAs regulate virus proliferation.关键词
草地贪夜蛾/Sf9细胞/苜蓿银纹夜蛾核多角体病毒/外泌体/microRNA/sfr-miR-1a-3pKey words
Spodoptera frugiperda/Sf9 cell/Autographa californica multiple nucleopolyhedrovirus/Exosome/microRNA/sfr-miR-1a-3p分类
生物科学引用本文复制引用
田伟彬,张以农,任菲菲,严寄铭,孙京臣..草地贪夜蛾Sf9细胞外泌体microRNA调控AcMNPV增殖的研究[J].华南农业大学学报,2025,46(2):133-140,8.基金项目
中国博士后科学基金(2022M721207) （2022M721207）
国家自然科学基金(32202749) （32202749）
广东省自然科学基金(2023A1515011658) （2023A1515011658）
