Department of Paediatrics, Prince of Wales Hospital, The Chinese University of Hong Kong, Shatin, Hong Kong; and.
Department of Surgery, Prince of Wales Hospital, The Chinese University of Hong Kong, Shatin, Hong Kong.
FASEB J. 2019 Apr;33(4):5143-5152. doi: 10.1096/fj.201801470R. Epub 2019 Jan 9.
The level of microRNA (miR)-431 was found to be markedly up-regulated in intestinal tissue of necrotizing enterocolitis (NEC). The objective of this study was to identify the target gene of miR-431 and to investigate the role of the miR-431-FOXA1 axis in the pathophysiology of NEC. The target gene of miR-431 was identified by in silico target prediction bioinformatics, luciferase assay, and Western blotting. Effects of miR-431 on downstream expression signals, cell proliferation, and apoptosis were investigated by overexpression in Caco-2 cells upon stimulation by LPS or lipoteichoic acid (LTA). FOXA1 was identified as the target gene of miR-431. Overexpression of miR-431 in Caco-2 cells significantly inhibited FOXA1, ESRRG, and HNF4A and activated IL-6, LGR5, NFKB2, PLA2G2A, PRKCZ, and TNF. IL-8 and - 10 were enhanced when costimulated with LPS or LTA. These potential downstream genes were also significantly dysregulated in primary NEC tissues compared with surgical-control tissues. Overexpression of miR-431 significantly decreased proliferation and increased apoptosis of Caco-2 cells. A proposed network of miR-431-FOXA1 interaction with LPS and LTA receptors demonstrates dysregulation of transcription factors, inflammatory mediators, epithelium tight junction regulators, and cell proliferation and apoptosis signals. The miR-431-FOXA1 axis could in part be responsible for the intensification of the inflammatory response in NEC tissues and contribute to the proinflammatory pathophysiology.-Wu, Y. Z., Chan, K. Y. Y., Leung, K. T., Lam, H. S., Tam, Y. H., Lee, K. H., Li, K., Ng, P. C. Dysregulation of miR-431 and target gene FOXA1 in intestinal tissues of infants with necrotizing enterocolitis.
miR-431 的水平在坏死性小肠结肠炎（NEC）的肠组织中明显上调。本研究旨在确定 miR-431 的靶基因，并探讨 miR-431-FOXA1 轴在 NEC 病理生理学中的作用。通过计算机预测生物信息学、荧光素酶测定和 Western blot 鉴定 miR-431 的靶基因。通过 LPS 或脂磷壁酸（LTA）刺激 Caco-2 细胞过表达 miR-431，研究 miR-431 对下游表达信号、细胞增殖和凋亡的影响。FOXA1 被鉴定为 miR-431 的靶基因。Caco-2 细胞中 miR-431 的过表达显著抑制 FOXA1、ESRRG 和 HNF4A，并激活 IL-6、LGR5、NFKB2、PLA2G2A、PRKCZ 和 TNF。当与 LPS 或 LTA 共刺激时，IL-8 和 -10 增强。与手术对照组织相比，这些潜在的下游基因在原发性 NEC 组织中也明显失调。miR-431 的过表达显著降低了 Caco-2 细胞的增殖并增加了其凋亡。miR-431-FOXA1 与 LPS 和 LTA 受体相互作用的网络表明转录因子、炎症介质、上皮紧密连接调节剂以及细胞增殖和凋亡信号的失调。miR-431-FOXA1 轴可能部分负责 NEC 组织中炎症反应的加剧，并有助于促炎病理生理学。-Wu，Y.Z.，Chan，K.Y.Y.，Leung，K.T.，Lam，H.S.，Tam，Y.H.，Lee，K.H.，Li，K.，Ng，P.C. miR-431 和靶基因 FOXA1 在坏死性小肠结肠炎婴儿肠道组织中的失调。
