Abstract：
Long non-coding RNA (lncRNA) is a novel emerging type of competitive endogenous RNA (ceRNA) that performs key functions in multiple biological processes. However, little is known about the roles of lncRNA under hypoxia stress in fish. Here, vascular endothelial growth factor-Aa (vegfaa) was cloned in rainbow trout (Oncorhynchus mykiss), with the complete cDNA sequence of 2914 bp, encoding 218 amino acids. The molecular weight of the protein was approximately 25.33 kDa, and contained PDGF and VEGF_C domains. Time-course and spatial expression patterns revealed that LOC110520012 was a key regulator of rainbow trout in response to hypoxia stress, and LOC110520012, miR-206-y and vegfaa exhibited a ceRNA regulatory relationship in liver, gill, muscle and rainbow trout liver cells treated with acute hypoxia. Subsequently, the targeting relationship of LOC110520012 and vegfaa with miR-206-y was confirmed by dual-luciferase reporter analysis, and overexpression of LOC110520012 mediated the inhibition of miR-206-y expression in rainbow trout liver cells, while the opposite results were obtained after LOC110520012 silencing with siRNA. We also proved that vegfaa was a target of miR-206-y in vitro and in vivo, and the vegfaa expression and anti-proliferative effect on rainbow trout liver cells regulated by miR-206-y mimics could be reversed by LOC110520012. These results suggested that LOC110520012 can positively regulate vegfaa expression by sponging miR-206-y under hypoxia stress in rainbow trout, which facilitate in-depth understanding of the molecular mechanisms of fish adaptation and tolerance to hypoxia.
摘要：
长链非编码RNA（lncRNA）是一种新型的竞争性内源性RNA（ceRNA），在多种生物过程中发挥关键功能。然而,lncRNA在鱼类缺氧胁迫下的作用知之甚少。这里,血管内皮生长因子-Aa（vegfaa）被克隆到虹鳟鱼（Oncorhynchusmykiss）中，具有2914bp的完整cDNA序列，编码218个氨基酸。蛋白质的分子量约为25.33kDa，并含有PDGF和VEGF_C结构域。时间过程和空间表达模式显示LOC110520012是虹鳟鱼响应低氧胁迫的关键调节因子,和LOC110520012，miR-206-y和vegfaa在肝脏中表现出ceRNA调控关系，吉尔,急性缺氧处理的肌肉和虹鳟鱼肝细胞。随后，通过双荧光素酶报告基因分析证实了LOC110520012和vegfaa与miR-206-y的靶向关系，LOC110520012的过表达介导miR-206-y在虹鳟鱼肝细胞中的表达抑制，而在用siRNA沉默LOC110520012后获得相反的结果。我们还证明vegfaa是miR-206-y在体外和体内的靶标，通过LOC110520012可以逆转miR-206-y模拟物调节的vegfaa对虹鳟鱼肝细胞的表达和抗增殖作用。这些结果表明，LOC110520012可以通过在低氧胁迫下海绵作用miR-206-y来正向调节虹鳟鱼vegfaa的表达，这有助于深入了解鱼类适应和耐缺氧的分子机制。
