广东海洋大学学报2025，Vol.45Issue(2)：43-51,9.DOI:10.3969/j.issn.1673-9159.2025.02.006
草鱼miR-34a在鱼淋巴囊肿病毒中国株感染过程中的调控机制
Regulatory Mechanism of Ctenopharyngodon idella miR-34a during Infection of Lymphocystis Disease Virus China
摘要
Abstract
[Objective]To explore the regulatory role of Ctenopharyngodon idella miR-34a(cid-miR-34a)on the replication of lymphocystis disease virus China(LCDV-cn),and reveal the regulatory mechanism of cid-miR-34a during the infection of LCDV-cn in grass carp ovary cells(GCO),and provide new evidence for clarifying the pathogenesis of LCDV-cn and treatment of lymphocystis disease.[Method]Cid-miR-34a was identified by stem-loop RT-PCR and sequencing.The temporal expression of cid-miR-34a during the infection of LCDV-cn in GCO cells was determined with qRT-PCR.Bioinformatic prediction and dual luciferase reporter gene assay were used to verify the regulatory factor lncRNA(long non-coding RNA)and the target gene of cid-miR-34a.Cid-miR-34a mimic was transfected to overexpress cid-miR-34a,and cid-miR-34a inhibitor was transfected to inhibit the expression of cid-miR-34a.The expression of cid-miR-34a,lncRNA,the target gene and the LCDV-cn mcp(major capsid protein)gene was detected by qRT-PCR.[Result]The length of cid-miR-34a is 22 bp,and it has strong conservation.During the infection of LCDV-cn in GCO cells,the expression level of cid-miR-34a increased gradually after infection,and decreased after reaching the peak at 72 hours(P<0.05).LncRNA LRP1 specifically bound to cid-miR-34a,and lncRNA LRP1 regulated the release of cid-miR-34a.Meanwhile,cid-miR-34a targeted to the gene lrp1(low density lipoprotein receptor-related protein 1),and regulated the expression of the gene lrp1.The lncRNA LRP1/cid-miR-34a/lrp1 axis was formed with lncRNA LRP1,cid-miR-34a and the target gene lrp1.After infection of LCDV-cn in GCO cells,the expression of lncRNA LRP1 showed an upward trend,which was positively corelated with the infection of LCDV-cn.After overexpression of cid-miR-34a,the expression of the target gene lrp1 decreased,and the expression of the LCDV-cn mcp gene and titers of LCDV-cn also decreased;after inhibition of the expression of cid-miR-34a,the expression of the gene lrp1 increased,and the expression of the LCDV-cn mcp gene and titers of LCDV-cn increased,too.[Conclusion]During the infection of LCDV-cn in GCO cells,the lncRNA LRP1/cid-miR-34a/lrp1 axis regulated the infection and replication of LCDV-cn.LncRNA LRP1 competitively bound to cid-miR-34a,and cid-miR-34a targeted to the gene lrp1 and inhibited its expression.LRP1 also played roles in promoting the replication of LCDV-cn and disease progression.Cid-miR-34a,lncRNA LRP1 and the target gene lrp1 were all potential targets for controlling the infection of LCDV-cn and treatment of lymphocystis disease.关键词
鱼淋巴囊肿病毒中国株/草鱼卵巢细胞/cid-miR-34a/长非编码RNA/低密度脂蛋白受体相关蛋白1Key words
lymphocystis disease virus China/grass carp ovary cells/cid-miR-34a/lncRNA/low density lipoprotein receptor-related protein 1分类
水产学引用本文复制引用
战盈瑾,周杰,臧辰禧,马嘉霖,闫秀英,简纪常..草鱼miR-34a在鱼淋巴囊肿病毒中国株感染过程中的调控机制[J].广东海洋大学学报,2025,45(2):43-51,9.基金项目
广东省科技特派员专项(GDKTP2021029800) （GDKTP2021029800）
