Yu ZeWen, Ren WenZhi, Wang Tian, Zhang WeiDi, Wang ChangJiang, Wang HaoQi, Gao Fei, Yuan Bao
Department of Laboratory Animals, College of Animal Sciences, Jilin University, 5333 Xi'an Road, Changchun 130062, China.
Animals (Basel). 2021 Feb 20;11(2):558. doi: 10.3390/ani11020558.
CircRNAs have been identified to be expressed differently and stably in numerous species and tissues, but their functions in growth hormone (GH) secretion are still largely unknown. In summary, we have revealed a circRNA-miRNA-mRNA network that may play a biological role in the rat pituitary gland. First, we verified the chromosome location information of circAgtpbp1 according to sequencing analysis. The circAgtpbp1 characteristics were authenticated through PCR, qRT-PCR, treating with RNase and fluorescent in situ hybridization (FISH). Second, we detected the expression pattern of circAgtpbp1 in the rat anterior pituitary by qRT-PCR. We also designed circAgtpbp1 siRNA and constructed overexpression plasmid to evaluate the effect of circAgtpbp1 function on GH secretion by qRT-PCR, ELISA and Western blot. CircAgtpbp1 is a stable, truly circular molecule. We found that circAgtpbp1 interacted with miR-543-5p and can regulate GH secretion in pituitary cells through a circAgtpbp1-miR-543-5p-GH axis. Overall, the evidence generated by our study suggests that circAgtpbp1 can act as a sponge of miR-543-5p to reduce the inhibitory effect of miR-543-5p on Gh1 and further promote GH secretion. These findings expand our existing knowledge on the mechanisms of hormone regulation in the pituitary gland.
已发现环状RNA(circRNAs)在众多物种和组织中存在差异表达且表达稳定,但其在生长激素(GH)分泌中的功能仍 largely未知。总之,我们揭示了一个可能在大鼠垂体中发挥生物学作用的circRNA- miRNA - mRNA网络。首先,我们根据测序分析验证了circAgtpbp1的染色体定位信息。通过PCR、qRT - PCR、用核糖核酸酶处理以及荧光原位杂交(FISH)对circAgtpbp1的特征进行了鉴定。其次,我们通过qRT - PCR检测了circAgtpbp1在大鼠垂体前叶中的表达模式。我们还设计了circAgtpbp1的小干扰RNA(siRNA)并构建了过表达质粒,通过qRT - PCR、酶联免疫吸附测定(ELISA)和蛋白质免疫印迹法(Western blot)评估circAgtpbp1功能对GH分泌的影响。circAgtpbp1是一个稳定的、真正的环状分子。我们发现circAgtpbp1与miR - 543 - 5p相互作用,并可通过circAgtpbp1 - miR - 543 - 5p - GH轴调节垂体细胞中的GH分泌。总体而言,我们的研究证据表明,circAgtpbp1可作为miR - 543 - 5p的海绵,以降低miR - 543 - 5p对Gh1的抑制作用,并进一步促进GH分泌。这些发现扩展了我们对垂体中激素调节机制的现有认识。