Yang Jie, Gong Yafan, Cai Jingzeng, Liu Qi, Zhang Ziwei
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, P. R. China.
College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, P. R. China; Key Laboratory of the Provincial Education Department of Heilongjiang for Common Animal Disease Prevention and Treatment, College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, P. R. China.
Mol Ther Nucleic Acids. 2019 Dec 6;18:1-15. doi: 10.1016/j.omtn.2019.08.003. Epub 2019 Aug 12.
Selenium deficiency has been proven to induce calcium disorders in the chicken heart. However, detailed regulatory mechanisms, e.g., the long noncoding RNA (lncRNA)-microRNA (miRNA)-mRNA regulatory axis, have not yet been described. Here, we point out lnc-2315, miR-1594, and Troponin T (TNNT2) based on the results of lncRNA and miRNA comparative genomics group analysis of Se-deficient chicken hearts compared with control hearts. We employed lnc-3215 and TNNT2 knockdown, miR-1594 knockdown, and overexpression models in the chicken embryos in vivo, and lnc-3215, miR-1594, and TNNT2 knockdown and overexpression models in cardiomyocytes in vitro. The dual-luciferase reporter assay and quantitative real-time PCR were used to confirm the relationships between miR-1594 and TNNT2, lnc-3215, and miR-1594 in cardiomyocytes. Our results revealed that TNNT2 suppression induced cardiac calcium overload in vivo and in vitro. miR-1594 activates cardiac calcium overload by targeting TNNT2. Moreover, we found that lnc-3215 regulates miR-1594, and thus influences the TNNT2 expression in vivo and in vitro; these conclusions were verified by gene knockdown in chicken embryos. Our present study revealed a novel regulatory model of a calcium program, which comprises lnc-3215, miR-1594, and TNNT2 in the chicken heart. Our conclusions may provide a feasible diagnostic tool for Se-deficient cardiomyocytes injury.
硒缺乏已被证明会诱发鸡心脏的钙紊乱。然而，详细的调控机制，如长链非编码RNA（lncRNA）-微小RNA（miRNA）-信使核糖核酸（mRNA）调控轴，尚未见报道。在此，我们基于与对照心脏相比的缺硒鸡心脏的lncRNA和miRNA比较基因组学组分析结果，指出了lnc-2315、miR-1594和肌钙蛋白T（TNNT2）。我们在体内鸡胚胎中采用了lnc-3215和TNNT2敲低、miR-1594敲低和过表达模型，以及在体外心肌细胞中采用了lnc-3215、miR-1594和TNNT2敲低和过表达模型。双荧光素酶报告基因检测和定量实时PCR用于确认心肌细胞中miR-1594与TNNT2、lnc-3215和miR-1594之间的关系。我们的结果显示，TNNT2抑制在体内和体外均诱导心脏钙超载。miR-1594通过靶向TNNT2激活心脏钙超载。此外，我们发现lnc-3215调节miR-1594，从而在体内和体外影响TNNT2的表达；这些结论通过鸡胚胎中的基因敲低得到了验证。我们目前的研究揭示了鸡心脏中由lnc-3215、miR-1594和TNNT2组成的钙程序的新调控模型。我们的结论可能为缺硒心肌细胞损伤提供一种可行的诊断工具。
