Department of Pediatrics, Heping Hospital Affiliated to Changzhi Medical College, No. 110, Yanan Sourth Road, Changzhi, 046000, Shanxi Province, China.
Department of Infectious Diseases, Heping Hospital Affiliated to Changzhi Medical College, Shanxi, China.
Genes Genomics. 2021 Aug;43(8):837-846. doi: 10.1007/s13258-021-01092-1. Epub 2021 Apr 17.
Asthma is a serious respiratory disease that affects the physical and mental health of children. Airway epithelial apoptosis concomitantly mediated by transforming growth factor-β1 (TGF-β1) is a crucial component of asthma pathogenesis. LncRNA growth Arrest Specific 5 (GAS5), microRNA-217 (miR-217) and Histone deacetylase 4 (HDAC4) shown a close relationship with TGF-β1-induced injury of airway epithelial. However, the mechanism underlying TGF-β1-induced injury of airway epithelial in asthma still needs to be investigated.
We aimed to investigate the effect and underlying mechanism of GAS5/miR-217/HDAC4 axis in TGF-β1-stimulated bronchial epithelial cells.
The levels of were detected by quantitative real-time polymerase chain reaction (RT-qPCR). All protein levels were determined by western blot. Cell viability and apoptosis rate were assessed by Methyl thiazolyl tetrazolium (MTT) and Flow cytometry, respectively. The targeting relationship between miR-217 and GAS5 or HDAC4 was examined with dual-luciferase reporter assay.
TGF-β1, GAS5, HDAC4 were up-regulated, while miR-217 was down-regulated in bronchial mucosal tissues of asthmatic children and TGF-β1-treated BEAS-2B cells. TGF-β1 could reduce cell viability and induce apoptosis, while these effects could be reversed by downregulation of GAS5 or HDAC4. Mechanically, GAS5 acted as a sponge for miR-217 to regulate the expression of HDAC4. Furthermore, overexpression of HDAC4 rescued the effects of GAS5 knockdown on viability and apoptosis of TGF-β1-induced BEAS-2B cells. GAS5 knockdown induced cell viability and hampered cell apoptosis in TGF-β1-stimulated BEAS-2B cells by regulating the miR-217/HDAC4 axis.
The lncRNA GAS5/miR-217/HDAC4 axis played an important role in regulating TGF-β1-induced bronchial epithelial cells injury, thus contributing to asthma.
哮喘是一种严重的呼吸道疾病，影响儿童的身心健康。转化生长因子-β1（TGF-β1）介导的气道上皮细胞凋亡是哮喘发病机制的重要组成部分。长链非编码 RNA（lncRNA）生长停滞特异性基因 5（GAS5）、微小 RNA-217（miR-217）和组蛋白去乙酰化酶 4（HDAC4）与 TGF-β1 诱导的气道上皮损伤密切相关。然而，TGF-β1 诱导的哮喘气道上皮损伤的机制仍需进一步研究。
探讨 GAS5/miR-217/HDAC4 轴在 TGF-β1 刺激的支气管上皮细胞中的作用及机制。
采用实时定量聚合酶链反应（RT-qPCR）检测水平。Western blot 法检测所有蛋白水平。噻唑蓝（MTT）比色法和流式细胞术分别评估细胞活力和凋亡率。双荧光素酶报告基因检测 miR-217 与 GAS5 或 HDAC4 的靶向关系。
哮喘患儿支气管黏膜组织和 TGF-β1 处理的 BEAS-2B 细胞中，TGF-β1、GAS5、HDAC4 上调，miR-217 下调。TGF-β1 可降低细胞活力并诱导细胞凋亡，下调 GAS5 或 HDAC4 可逆转这些作用。机制上，GAS5 作为 miR-217 的海绵体调节 HDAC4 的表达。此外，过表达 HDAC4 可挽救 GAS5 敲低对 TGF-β1 诱导的 BEAS-2B 细胞活力和凋亡的影响。GAS5 敲低通过调节 miR-217/HDAC4 轴在 TGF-β1 刺激的 BEAS-2B 细胞中诱导细胞活力并阻碍细胞凋亡。
lncRNA GAS5/miR-217/HDAC4 轴在调节 TGF-β1 诱导的支气管上皮细胞损伤中起重要作用，从而促进哮喘的发生。
