胡海浩,黄鉴涛,马嘉霖,杨硕,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】在鱼淋巴囊肿中国病毒株（LCDV-cn）感染草鱼卵巢细胞系（GCO）过程中，分析cid-miR-146a的表达特性，探索cid-miR-146a的调控作用。【方法】采用颈环RT-PCR方法，从LCDV-cn感染的GCO细胞中获得cidmiR-146a；在LCDV-cn感染GCO过程中，用定量PCR方法获取cid-miR-146a的表达变化；用生物信息学方法预测和双荧光素酶系统验证cid-miR-146a的靶基因；转染cid-miR-146a mimic和cid-miR-146a inhibitor对cid-miR-146a进行上调和下调，用定量PCR检测cid-miR-146a、靶基因Flt1和LCDV-cn mcp基因在GCO中的表达。【结果】LCDV-cn感染GCO后3～6 d细胞聚集形成“疤痕”，之后细胞逐渐脱落、裂解，呈现空洞。cid-miR-146a的长度为23 bp，在LCDV-cn感染GCO过程中，cid-miR-146a的表达先上升（72 h前）后下降（72 h后）。用不同生物信息学方法共同预测到cid-miR-146a的靶基因为Flt1，在双荧光素酶系统验证实验中，cid-miR-146a靶向Flt1重组载体后荧光素酶活性降低（P<0.05），证实Flt1为cid-miR-146a的靶基因。对cid-miR-146a进行上调下调后，GCO中cid-miR-146a的表达差异显著（P <0.05）。在cid-miR-146a上调后，靶基因Flt1的表达显著下降（P <0.05）,LCDV-cn mcp基因的表达显著上升（P<0.05）；在cid-miR-146a下调后，靶基因Flt1的表达显著上升（P<0.05）,LCDV-cn mcp基因的表达显著下降（P<0.05）。【结论】在LCDV-cn感染GCO过程中，CPE的变化与cid-miR-146a的表达变化时间点呈正相关。cid-miR-146a负调控其靶基因Flt1的表达，并对LCDV-cn的复制起着正调控作用。生物信息学方法预测说明cid-miR-146a参与调控的信号通路与免疫和肿瘤发生相关。

关键词(KeyWords)： 鱼淋巴囊肿病毒中国株;草鱼卵巢细胞系;草鱼miR-146a;Fms相关酪氨酸激酶1;定量PCR

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金（31602199）;; 广东省企业科技特派员专项（GDKTP2021029800）

作者(Author): 胡海浩,黄鉴涛,马嘉霖,杨硕,闫秀英,简纪常

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