Department of Gastrointestinal Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Department of Gastrointestinal Surgery II, Renmin Hospital of Wuhan University, Wuhan, 430060, China.
Pediatr Res. 2019 Oct;86(4):460-470. doi: 10.1038/s41390-019-0324-9. Epub 2019 Mar 1.
The miR-214 has been reported to be associated with various diseases, but its involvement in the pathophysiology of Hirschsprung disease (HSCR) is almost completely unexplored.
In our study, we conducted a series of experiments to unravel the biological role of miR-214 in the pathophysiology of HSCR. qRT-PCR and western blotting were utilized to investigate the relative expression levels of miR-214, mRNAs, and proteins of related genes in colon tissues from 20 controls without HSCR and 24 patients with HSCR. The potential biological role of miR-214 in two cell lines (SKN-SH and SH-SY5Y) was assessed using the CCK8 assay, EdU staining, transwell assay, and flow cytometry. The dual-luciferase reporter assay was used to confirm PLAGL2 as a common target gene of miR-214.
All results suggested that miR-214 is upregulated in HSCR tissue samples compared with controls. Additionally, we found that miR-214 could inhibit cell proliferation and migration by directly downregulating the expression of PLAGL2, and the extent of the miR-214-mediated inhibitory effects could be rescued by a PLAGL2 overexpression plasmid.
Our results revealed that miR-214 is indeed involved in the pathophysiology of HSCR and suppresses cell proliferation and migration by directly downregulating PLAGL2 in cell models.
miR-214 与多种疾病相关,但它在先天性巨结肠病(HSCR)病理生理学中的作用几乎完全未知。
在本研究中,我们进行了一系列实验,以阐明 miR-214 在 HSCR 病理生理学中的生物学作用。我们利用 qRT-PCR 和 Western blot 检测了 20 例无 HSCR 对照和 24 例 HSCR 患者结肠组织中 miR-214、mRNA 和相关基因蛋白的相对表达水平。利用 CCK8 检测、EdU 染色、Transwell 检测和流式细胞术评估了 miR-214 在两种细胞系(SKN-SH 和 SH-SY5Y)中的潜在生物学作用。双荧光素酶报告基因检测用于证实 PLAGL2 是 miR-214 的共同靶基因。
所有结果均表明,与对照组相比,miR-214 在 HSCR 组织样本中上调。此外,我们发现 miR-214 可通过直接下调 PLAGL2 的表达来抑制细胞增殖和迁移,而 PLAGL2 过表达质粒可挽救 miR-214 介导的抑制作用。
我们的研究结果表明,miR-214 确实参与了 HSCR 的病理生理学过程,并通过直接下调 PLAGL2 来抑制细胞增殖和迁移。