战盈瑾,周杰,臧辰禧,马嘉霖,闫秀英,简纪常

• 1:广东海洋大学水产学院/广东省水产动物病害防控与健康养殖重点实验室/水产经济动物病害控制广东普通高校重点实验室

摘要(Abstract)：

【目的】探索草鱼（Ctenopharyngodonidella）miR-34a(cid-miR-34a)对鱼淋巴囊肿病毒中国株（Lymphocystis disease virus China,LCDV-cn）复制的调控作用，揭示cid-miR-34a在LCDV-cn感染草鱼卵巢细胞（Grass carp ovary cells,GCO）过程中的调控机制，为LCDV-cn致病机制解析和淋巴囊肿病治疗提供新的依据。【方法】应用茎环RT-PCR获得cid-miR-34a，并测序鉴定；采用定量PCR(qRT-PCR)测定cid-miR-34a在LCDV-cn感染GCO过程中的时序表达；通过生物信息学预测和双荧光素酶报告基因实验，确定cid-miR-34a的调控因子长非编码RNA(Long non-coding RNA,lncRNA)及其靶基因；转染cid-miR-34a mimic过表达cid-miR-34a，转染cid-miR-34a inhibitor抑制cid-miR-34a表达，并用qRT-PCR检测cid-miR-34a、lncRNA、靶基因和LCDV-cn主要衣壳蛋白基因mcp的表达。【结果】cid-miR-34a长度为22 bp，保守性强。在LCDV-cn感染GCO过程中，cid-miR-34a的表达量逐渐升高，在感染72 h达峰值后下降。lncRNA LRP1靶向结合cid-miR-34a，并调控cid-miR-34a的释放，且cid-miR-34a靶向调控低密度脂蛋白受体相关蛋白1基因lrp1的表达，lncRNA LRP1、cid-miR-34a和靶基因lrp1形成lncRNA LRP1/cid-miR-34a/lrp1轴。LCDV-cn感染GCO后，lncRNA LRP1的表达量呈上升趋势，与LCDV-cn的感染呈正相关。过表达cid-miR-34a后，lrp1表达量降低，LCDV-cn mcp表达和病毒滴度也降低；抑制cid-miR-34a表达后，lrp1表达量升高，LCDV-cn mcp表达和病毒滴度也升高。【结论】在LCDV-cn感染GCO过程中，lncRNA LRP1/cid-miR-34a/lrp1轴调控着病毒感染和复制。lncRNA LRP1竞争性结合cid-miR-34a,cid-miR-34a靶向抑制lrp1的表达，LRP1又发挥着促进LCDV-cn复制和疾病发展的作用。cid-miR-34a、lncRNA LRP1和lrp1均是抑制LCDV-cn感染和治疗淋巴囊肿病的潜在靶标。

关键词(KeyWords)： 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1

Abstract:

Keywords:

基金项目(Foundation): 广东省科技特派员专项（GDKTP2021029800）

作者(Author): 战盈瑾,周杰,臧辰禧,马嘉霖,闫秀英,简纪常

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