Department of General Surgery, Hunan Children's Hospital, Changsha, P.R. China.
Department of General Surgery, Hunan Children's Hospital, Changsha, P.R. China;
Acta Neurobiol Exp (Wars). 2022;82(3):336-346. doi: 10.55782/ane-2022-032.
Hirschsprung's disease (HSCR) is an intestinal disease caused by defects in neural crest cell migration, proliferation, differentiation, and survival. Many reports have proposed that miRNA dysregulation is related to the occurrence of HSCR. However, the roles and mechanisms of miRNAs have not been thoroughly studied. The levels of miR‑92a and KLF4 were examined using qRT‑PCR and immunohistochemistry, respectively. Cell viability, migration and apoptosis were evaluated by MTT, Transwell and flow cytometry assays, respectively. A dual‑luciferase reporter assay was employed to verify the binding relationship between miR‑92a and KLF4. Levels of PI3K/AKT signals were further determined by western blot assay. Herein, elevated expression of miR‑92a and reduced expression of KLF4 were found in HSCR tissues, and their expression patterns were negatively correlated. Overexpression of miR‑92a inhibited cell viability and migration but enhanced cell apoptosis. However, overexpression of KLF4 had the opposite effects. Mechanistically, KLF4 was a target of miR‑92a and it negatively affected biological functions by activating PI3K/AKT signaling. These results proved that miR‑92a inhibited the proliferation and metastasis of nerve cells by regulating the KLF4/PI3K/AKT axis.
先天性巨结肠症（HSCR）是一种由神经嵴细胞迁移、增殖、分化和存活缺陷引起的肠道疾病。许多报道提出 miRNA 失调与 HSCR 的发生有关。然而，miRNA 的作用和机制尚未得到彻底研究。采用 qRT-PCR 和免疫组织化学分别检测 miR-92a 和 KLF4 的水平。通过 MTT、Transwell 和流式细胞术分别评估细胞活力、迁移和凋亡。双荧光素酶报告基因实验验证 miR-92a 与 KLF4 的结合关系。通过 Western blot 进一步确定 PI3K/AKT 信号水平。在此，发现 HSCR 组织中 miR-92a 表达升高，KLF4 表达降低，且表达模式呈负相关。miR-92a 的过表达抑制细胞活力和迁移，但促进细胞凋亡。然而，KLF4 的过表达则具有相反的效果。机制上，KLF4 是 miR-92a 的靶基因，通过激活 PI3K/AKT 信号通路来负调控生物学功能。这些结果证明，miR-92a 通过调节 KLF4/PI3K/AKT 轴抑制神经细胞的增殖和迁移。
