The Cleft Lip and Palate Treatment Center, Second Affiliated Hospital of Shantou University Medical College, Shantou 515041, China.
Department of Plastic and Aesthetic Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Exp Biol Med (Maywood). 2023 Jul;248(13):1124-1133. doi: 10.1177/15353702231182215. Epub 2023 Aug 1.
MicroRNAs (miRNAs) have been identified as crucial modulators of gene expression and to play a role in palatogenesis. The aim of this study was to explore the potential role and regulatory mechanisms of miRNAs during palatogenesis. RNA-sequencing was performed to compare the RNA expression profiles of mouse embryonic palatal shelf (MEPS) tissue between an all-trans retinoic acid (ATRA)-induced group and control group, followed by reverse transcription-quantitative polymerase chain reaction for validation, demonstrating upregulated expression of miRNA-470-5p and downregulated expression of in the ATRA-induced group. The specific binding sites of miRNA-470-5p that potentially govern expression were predicted by miRanda and TargetScan. The relationship between miRNA-470-5p and was validated in HEK293T cells by luciferase reporter assays, confirming that miR-470-5p acts directly on the 3'-untranslated region. mRNA and FGFR1 protein levels were markedly downregulated in MEPS epithelial cells over-expressing miRNA-470-5p. Functional experiments with CCK-8, cell colony formation, and 5-ethynyl-2'-deoxyuridine (EdU) staining assays revealed that upregulated miRNA-470-5p expression could inhibit the epithelial-mesenchymal transition (EMT) of MEPS epithelial cells by targeting . These findings provide a new molecular mechanism of cleft palate formation, which can inform the development of new treatment and/or prevention targets.
微小 RNA(miRNA)已被鉴定为基因表达的关键调控因子,并在腭形成中发挥作用。本研究旨在探讨 miRNA 在腭形成过程中的潜在作用和调控机制。通过 RNA 测序比较全反式视黄酸(ATRA)诱导组和对照组小鼠胚胎腭突(MEPS)组织的 RNA 表达谱,随后进行逆转录定量聚合酶链反应验证,结果显示 ATRA 诱导组 miRNA-470-5p 表达上调,而 表达下调。通过 miRanda 和 TargetScan 预测了 miRNA-470-5p 潜在调控 表达的特异性结合位点。通过荧光素酶报告基因实验在 HEK293T 细胞中验证了 miRNA-470-5p 与 的关系,证实了 miR-470-5p 直接作用于 的 3'-非翻译区。在过表达 miRNA-470-5p 的 MEPS 上皮细胞中, mRNA 和 FGFR1 蛋白水平明显下调。CCK-8、细胞集落形成和 5-乙炔基-2'-脱氧尿苷(EdU)染色功能实验表明,上调 miRNA-470-5p 表达可通过靶向 抑制 MEPS 上皮细胞的上皮-间充质转化(EMT)。这些发现为腭裂形成提供了一个新的分子机制,可为新的治疗和/或预防靶点的开发提供信息。