BACKGROUND Hirschsprung's disease (HSCR) is characterized by missing of enteric neurons in the terminal areas of the whole gut, which is causally related to poor proliferation of enteric neural crest cells (ENCCs). Our aim is to explore how miR-431-5p interacts with its target gene in regulation of proliferation of ENCCs in HSCR. METHODS Mouse model of HSCR was established by Benzalkonium chloride (BAC) treatment. Quantitative Real-time PCR and western blotting were performed to determine the miR-431-5p and the LRSAM1 expression in colon tissues of the HSCR group (n = 8) and the control group (n = 8) and in ENCCs isolated from colon tissues. CCK-8 assay was performed to detect the proliferation of ENCCs of HSCR. ENCCs after transfection with miR-431-5p mimics or miR-431-5p inhibitor. Luciferase reporter assay was conducted to clarify the connections between miR-431-5p and LRSAM1. RESULTS Upregulation of miR-431-5p and downregulation of LRSAM1 were found in ENCCs of HSCR. Downregulation of miR-431-5p could promote cell proliferation of ENCCs. LRSAM1 was proved to be the target gene of miR-431-5p by luciferase reporter assay. Moreover, proliferation of ENCCs was increased in the miR-431-5p inhibitor group and was suppressed after knocking down LRSAM1. CONCLUSION Downregulation of miR-431-5p promoted proliferation of ENCCs via targeting LRSAM1, which provides an innovative and candidate target for treatment of HSCR.

中文翻译：

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microRNA-431-5p的下调通过靶向LRSAM1在Hirschsprung病中促进肠神经rest细胞增殖。

背景技术高发性肺病（HSCR）的特征在于整个肠末端区域中的肠神经元缺失，这与肠神经c细胞（ENCC）的增殖不良有关。我们的目的是探索miR-431-5p如何与其靶基因相互作用，调节HSCR中ENCC的增殖。方法采用苯扎氯铵（BAC）处理建立HSCR小鼠模型。进行实时荧光定量PCR和蛋白质印迹分析，以确定HSCR组（n = 8）和对照组（n = 8）的结肠组织以及从结肠组织分离的ENCC中的miR-431-5p和LRSAM1表达。进行CCK-8测定以检测HSCR的ENCC的增殖。用miR-431-5p模拟物或miR-431-5p抑制剂转染后的ENCC。进行荧光素酶报告基因分析以阐明miR-431-5p和LRSAM1之间的联系。结果在HSCR的ENCC中发现miR-431-5p的上调和LRSAM1的下调。miR-431-5p的下调可能促进ENCC的细胞增殖。萤光素酶报告基因测定证实LRSAM1是miR-431-5p的靶基因。此外，在miR-431-5p抑制剂组中ENCC的增殖增加，并在敲低LRSAM1后被抑制。结论miR-431-5p的下调通过靶向LRSAM1促进了ENCC的增殖，这为治疗HSCR提供了创新的候选靶点。萤光素酶报告基因测定证实LRSAM1是miR-431-5p的靶基因。此外，在miR-431-5p抑制剂组中ENCC的增殖增加，并在敲低LRSAM1后被抑制。结论miR-431-5p的下调通过靶向LRSAM1促进了ENCC的增殖，这为治疗HSCR提供了创新的候选靶点。萤光素酶报告基因测定证实LRSAM1是miR-431-5p的靶基因。此外，在miR-431-5p抑制剂组中ENCC的增殖增加，并在敲低LRSAM1后被抑制。结论miR-431-5p的下调通过靶向LRSAM1促进了ENCC的增殖，这为治疗HSCR提供了一个创新的候选靶标。