School of Marine Sciences, Ningbo University, PR China.
School of Marine Sciences, Ningbo University, PR China.
Fish Shellfish Immunol. 2017 Oct;69:211-217. doi: 10.1016/j.fsi.2017.08.033. Epub 2017 Aug 30.
miR-92a, a well-documented oncogene, was previously found to be differentially expressed in diseased sea cucumber Apostichopus japonicus by high-throughput sequencing. In this study, we identified Aj14-3-3ζ as a novel target of miR-92a in this species and investigated their regulatory roles in vivo. The negative expression profiles between miR-92a and Aj14-3-3ζ protein were detected in both LPS-exposed primary coelomocytes and Vibrio splendidus-challenged sea cucumbers. Over-expression of miR-92a by injection of miR-92a agomir significantly depressed the mRNA and protein expression of Aj14-3-3ζ and promoted coelomocytes apoptosis with 5.04-fold increase in vivo, which was consistent with those from siRNA-mediated Aj14-3-3ζ knockdown assay. In contrast, miR-92a antagomir significantly elevated the mRNA and protein expression of Aj14-3-3ζ and decreased coelomocytes apoptosis. Taken together, our result confirmed that miR-92a is involved in apoptotic signaling pathway regulation perhaps via targeting Aj14-3-3ζ in sea cucumbers, which will enhance our understanding of miR-92a regulatory roles in sea cucumber pathogenesis.
miR-92a 是一种已被充分研究的致癌基因，之前通过高通量测序发现其在患病海参刺参中差异表达。在本研究中，我们鉴定出 Aj14-3-3ζ 是该物种中 miR-92a 的一个新靶基因，并研究了它们在体内的调节作用。在 LPS 暴露的原腔细胞和灿烂弧菌攻毒的海参中，miR-92a 和 Aj14-3-3ζ 蛋白之间的负表达谱被检测到。通过注射 miR-92a agomir 过表达 miR-92a 显著抑制 Aj14-3-3ζ 的 mRNA 和蛋白表达，并促进体内原腔细胞凋亡，其凋亡率增加了 5.04 倍，与 siRNA 介导的 Aj14-3-3ζ 敲低实验结果一致。相反，miR-92a antagomir 显著提高了 Aj14-3-3ζ 的 mRNA 和蛋白表达，并降低了原腔细胞凋亡。总之，我们的结果证实，miR-92a 通过靶向海参中的 Aj14-3-3ζ 参与凋亡信号通路的调节，这将增强我们对 miR-92a 在海参发病机制中调节作用的理解。
