Pan Kaiyu, Jiang Xiaoguang, Hu Xiaohong, Zhan Jianhua, Zhang Chengyue
Department of Pediatrics, Xiaoshan Affiliated Hospital of Wenzhou Medical University, Hangzhou, Zhejiang, China.
Department of Pediatrics, Xiaoshan District Hospital of Traditional Chinese Medicine and Orthopedics, Hangzhou, Zhejiang, China.
J Pediatr Endocrinol Metab. 2024 Oct 15;37(12):1054-1065. doi: 10.1515/jpem-2024-0449. Print 2024 Dec 17.
Growth hormone deficiency (GHD) diagnosis still lacks a gold standard or ideal diagnostic marker. Unlike other epigenetic mechanisms, non-coding RNAs regulate post-transcriptional levels. The information on non-coding RNAs in the field of GHD is limited. Therefore, this study aimed to explore the role of hsa_circ_0002473 as a competitive endogenous RNA for has-miR-4645-3p in attenuating the inhibitory effect of has-miR-4645-3p on SSTR2. In this study, we screened three significantly expressed circular RNAs (circRNAs) in five children with GHD, and selected the highest expressed hsa_circ_0002473 as the study object, and screened has-miR-4645-3p, which is the most likely to bind to hsa_circ_0002473, according to the microRNA (miRNA)-circRNA regulatory network, to study the role and mechanism of has-miR-4645-3p as a competitive endogenous RNA of has-miR-4645-3p on GH3 cells. Somatostatin receptor 2 (SSTR2) inhibits GH3 cell proliferation, and miRNA binding to SSTR2 inhibits the latter expression. Both bioinformatics and dual-luciferase reporter analyses showed targeting relationships between hsa_circ_0002473 and has-miR-4645-3p and between has-miR-4645-3p and SSTR2. We constructed the hsa_circ_0002473/has-miR-4645-3p axis and transfected it into GH3 cells and found that overexpression of hsa_circ_0002473 inhibited the proliferation and growth hormone (GH) secretion of GH3 cells, and that hsa-miR-4645-3p promoted the proliferation and GH secretion of GH3 cells by targeting SSTR2. Co-culture revealed that the inhibitory effect of hsa_circ_0002473 was reversed by has-miR-4645-3p. In conclusion, our findings suggest that hsa_circ_0002473 can act as a competitive endogenous RNA for has-miR-4645-3p to regulate GH3 cell proliferation and secretion by targeting SSTR2.
生长激素缺乏症（GHD）的诊断仍缺乏金标准或理想的诊断标志物。与其他表观遗传机制不同，非编码RNA调节转录后水平。GHD领域中关于非编码RNA的信息有限。因此，本研究旨在探讨hsa_circ_0002473作为has-miR-4645-3p的竞争性内源性RNA在减弱has-miR-4645-3p对SSTR2的抑制作用中的作用。在本研究中，我们在5例GHD患儿中筛选出3种显著表达的环状RNA（circRNA），并选择表达量最高的hsa_circ_0002473作为研究对象，根据微小RNA（miRNA）-circRNA调控网络筛选出最有可能与hsa_circ_0002473结合的has-miR-4645-3p，以研究has-miR-4645-3p作为hsa_circ_0002473的竞争性内源性RNA对GH3细胞的作用及机制。生长抑素受体2（SSTR2）抑制GH3细胞增殖，miRNA与SSTR2结合会抑制后者的表达。生物信息学分析和双荧光素酶报告基因分析均显示hsa_circ_0002473与has-miR-4645-3p之间以及has-miR-4645-3p与SSTR2之间存在靶向关系。我们构建了hsa_circ_0002473/has-miR-4645-3p轴并将其转染到GH3细胞中，发现hsa_circ_0002473的过表达抑制了GH3细胞的增殖和生长激素（GH）分泌，而hsa-miR-4645-3p通过靶向SSTR2促进了GH3细胞的增殖和GH分泌。共培养显示hsa_circ_0002473的抑制作用被has-miR-4645-3p逆转。总之，我们的研究结果表明，hsa_circ_0002473可以作为has-miR-4645-3p的竞争性内源性RNA，通过靶向SSTR2来调节GH3细胞的增殖和分泌。
