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微小RNA-155-5p通过靶向组蛋白去乙酰化酶1调控鼻黏膜上皮细胞上皮间质转化的实验研究

Experimental study of microRNA-155-5p regulating epithelial mesenchymal transformation of nasal mucosal epithelial cells by targeting histone deacetylase 1

    摘要
  • 摘要:
    目的 观察微小RNA（miR）-155-5p对人鼻黏膜上皮细胞（HNEPC）上皮间质转化（EMT）的影响及其与组蛋白去乙酰化酶1（SIRT1）的靶向关系。
    方法 选取2019年5月—2021年12月行鼻内镜手术治疗的患者50例为研究对象。根据是否合并鼻息肉分为慢性鼻窦炎伴鼻息肉（CRSwNP）组36例和慢性鼻窦炎不伴鼻息肉（CRSsNP）组14例，另选取同期行鼻中隔偏曲手术治疗的20例鼻中隔偏曲患者为对照组。将miR-155-5p mimics、miR-155-5p inhibitor、miR-NC及Vector质粒转染至细胞，分为miR-155-5p组、miR-155-5p inhibitor组、miR-NC组和Vector组。实时荧光定量聚合酶链反应（RT-qPCR）检测细胞、组织miR-155-5p表达水平；Western blot、免疫荧光检测组织、细胞SIRT1、E-钙黏蛋白（E-cadherin）、波形蛋白（Vimentin）、α-平滑肌肌动蛋白（α-SMA）、纤黏蛋白（Fibronectin）表达；生物信息学工具、荧光素酶报告实验分析miR-155-5p与SIRT1的靶向关系。
    结果 CRSwNP组鼻腔黏膜组织miR-155-5p表达水平高于CRSsNP组及对照组，CRSsNP组鼻腔黏膜组织miR-155-5p表达水平高于对照组，差异有统计学意义（P < 0.01）。CRSwNP组鼻腔黏膜组织E-cadherin、SIRT1表达水平低于CRSsNP组及对照组，CRSsNP组鼻腔黏膜组织E-cadherin、SIRT1表达水平低于对照组，差异有统计学意义（P < 0.05）。CRSwNP组鼻腔黏膜组织Vimentin、α-SMA、Fibronectin表达水平高于CRSsNP组及对照组，CRSsNP组鼻腔黏膜组织Vimentin、α-SMA、Fibronectin表达水平高于对照组，差异有统计学意义（P < 0.05）。上调miR-155-5p促进了HNEPC细胞EMT，沉默miR-155-5p抑制了HNEPC细胞EMT。miR-155-5p负性靶向调控SIRT1。
    结论 miR-155-5p、EMT相关标记物在CRSwNP中表达升高，上调miR-155-5p可通过负性靶向调控SIRT1促进HNEPC的EMT过程。

     

    Abstract:
    Objective To observe the effect of microRNA(miR)-155-5p on epithelial mesenchymal transition (EMT) of human nasal epithelial cells (HNEPC) and its targeting relationship with histone deacetylase 1 (SIRT1).
    Methods A total of 50 patients who underwent endoscopic nasal surgery from May 2019 to December 2021 were selected as the research objects. According to whether nasal polyps were complicated or not, the patients were divided into chronic sinusitis with nasal polyps (CRSwNP) group (n=36) and chronic sinusitis without nasal polyps (CRSsNP) group (n=14). At the same time, 20 patients with nasal septum deviation who underwent nasal septum deviation surgery were selected as the control group. MiR-155-5p mimics, miR-155-5p inhibitor, miR-NC and vector plasmids were transfected into cells and divided into miR-155-5p group, miR-155-5p inhibitor group, miR-NC group and Vector group. Real time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression level of miR-155-5p in cells and tissues; western blot and immunofluorescence were used to detect tissue and cell SIRT1, E-cadherin, vimentin α smooth muscle actin (α-SMA), fibronectin expression; the bioinformatics tools and luciferase report experiment were used to analyze the targeting relationship between miR-155-5p and SIRT1.
    Results The expression level of miR-155-5p in nasal mucosa of the CRSwNP group was significantly higher than that of the CRSsNP group and control group, and the expression level of miR-155-5p in nasal mucosa of the CRSsNP group was significantly higher than that of the control group (P < 0.01). The expression levels of E-cadherin and SIRT1 in nasal mucosa of the CRSwNP group were significantly lower than those of the CRSsNP group and control group, and the expression levels of E-cadherin and SIRT1 in nasal mucosa of the CRSsNP group were significantly lower than those of the control group (P < 0.05). The expression levels of Vimentin, α-SMA and Fibronectin innasal mucosa of the CRSwNP group were significantly higher than those in the CRSsNP group and control group, and the expression levels of Vimentin, α-SMA and Fibronectin in nasal mucosa of the CRSsNP group were significantly higher than those in the control group (P < 0.05). Up-regulation of miR-155-5p promoted the EMT process of HNEPC cells, while silencing miR-155-5p inhibited the EMT process of HNEPC cells. MiR-155-5p negatively targets SIRT1.
    Conclusion The expressions of miR-155-5p and EMT related markers are increased in CRSwNP. Up-regulation of miR-155-5p can promote the EMT process of HNEPC by negatively targeting SIRT1.

     

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