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RASSF1‐AS1, an antisense lncRNA of RASSF1A, inhibits the translation of RASSF1A to exacerbate cardiac fibrosis in mice

机译:Rassf1-AS1,RASSF1A的反义LNCRNA,抑制RASSF1A在小鼠中加剧心脏纤维化的翻译

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Abstract Cardiac fibrosis is associated with various cardiovascular diseases and can eventually lead to heart failure. Dysregulation of long non‐coding RNAs (lncRNAs) are recognized as one of the key mechanisms of cardiac diseases. However, the roles and underlying mechanisms of lncRNAs in cardiac fibrosis have not been explicitly defined. Here, we investigated the role of an antisense (AS) lncRNA from the Ras association domain‐containing protein 1 isoform A (RASSF1A) gene locus, named RASSF1‐AS1, in the development of cardiac fibrosis. Cardiac fibrosis mouse model was established by isoproterenol injection. We found that RASSF1A protein was downregulated, whereas RASSF1‐AS1 was markedly upregulated during cardiac fibrosis. Overexpression and knockdown of mouse primary cardiac fibroblasts showed that RASSF1‐AS1 negatively regulated RASSF1A expression at the post‐transcriptional level. According to the landscape analysis and sense‐AS binding evaluation, RASSF1‐AS1 partially overlaps with RASSF1A messenger RNA (mRNA) at the exon2 region. RNA pull‐down and luciferase activity assays confirmed that RASSF1‐AS1 directly bound to RASSF1A mRNA and suppressed its translation. Furthermore, wild‐type RASSF1‐AS1 had a promoting effect on nuclear factor‐κB activation and cardiac fibrosis, but mutated RASSF1‐AS1, in which the binding region was deleted, had no effect. In conclusion, RASSF1‐AS1 inhibits the translation of RASSF1A to exacerbate cardiac fibrosis in mice, indicating a potential application of RASSF1‐AS1 as a therapy target for cardiac fibrosis.
机译:摘要心肌纤维化与各种心血管疾病有关,最终可能导致心力衰竭。长期非编码RNA(LNCRNA)的失调被认为是心脏病的关键机制之一。然而,没有明确定义LNCRNA在心肌纤维化中的角色和潜在机制。在这里,我们研究了反义(AS)LNCRNA从RAS关联结构域的蛋白质1同种型A(RASSF1A)基因座,命名为Rassf1-AS1的作用,在心脏纤维化的发展中。通过异丙肾镜注射建立心脏纤维化小鼠模型。我们发现rassf1a蛋白被下调,而rassf1-as1在心肌纤维化期间显着上调。对小鼠原发性心脏成纤维细胞的过度表达和敲低显示RASSF1-AS1在转录后水平下对RASSF1A表达产生负面调节。根据景观分析和感应 - 作为结合评估,RASSF1-AS1在EXON2区域的RASSF1A信使RNA(mRNA)部分重叠。 RNA下拉和荧光素酶活性测定证实,RASSF1-AS1直接与RASSF1A mRNA结合并抑制其翻译。此外,野生型RASSF1-AS1对核因子-κB活化和心肌纤维化具有促进作用,但突变的RASSF1-AS1,其中缺失结合区域无效。总之,Rassf1-AS1抑制RASSF1a在小鼠中加剧心肌纤维化的翻译,表明RASSF1-AS1作为心肌纤维化治疗靶标。

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