GATA1在支气管哮喘中的表达及对ORMDL3基因转录调控的影响

陈虎; 李娇娇; 袁越; 金蕊

doi:10.7499/j.issn.1008-8830.2408143

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中国当代儿科杂志 ›› 2025, Vol. 27 ›› Issue (2) : 212-218. DOI: 10.7499/j.issn.1008-8830.2408143

论著·实验研究

GATA1在支气管哮喘中的表达及对ORMDL3基因转录调控的影响

陈虎^1,2, 李娇娇¹, 袁越¹, 金蕊¹

作者信息 +

Expression of GATA1 in bronchial asthma and its effect on the transcription regulation of the ORMDL3 gene

CHEN Hu, LI Jiao-Jiao, YUAN Yue, JIN Rui

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文章历史 +

摘要

目的研究转录因子GATA1在哮喘中的表达及对支气管哮喘（简称哮喘）易感基因血清类黏蛋白1样蛋白3（orosomucoid 1-like protein 3, ORMDL3）表达水平的影响及分子机制研究。方法在基因表达综合（Gene Expression Omnibus, GEO）数据库中纳入中度哮喘组28例，重度哮喘组46例，正常对照组12例，分析各组GATA1和ORMDL3 mRNA表达水平及相关性。将pGL-185/58质粒分别与GATA1基因siRNA（si-GATA1组）、siRNA阴性对照（si-control）（si-control组）共转染BEAS-2B细胞。通过生物信息学方法预测ORMDL3基因启动子区域GATA1结合位点，采用双萤光素酶报告基因系统检测ORMDL3基因启动子活性，实时荧光定量PCR技术和蛋白质印迹法检测GATA1、ORMDL3 mRNA及蛋白表达水平，染色质免疫沉淀实验检测GATA1是否与ORMDL3启动子区结合。结果重度哮喘组GATA1、ORMDL3 mRNA表达水平较正常对照组明显升高（P<0.001）；中、重度哮喘组GATA1 mRNA与ORMDL3 mRNA表达水平呈正相关（分别r=0.636、0.341，均P<0.05）。BEAS-2B细胞中，双萤光素酶报告基因检测结果显示，si-GATA1组ORMDL3启动子萤光素酶活性、ORMDL3 mRNA及蛋白表达水平较si-control组降低（P<0.05）。染色质免疫沉淀实验结果显示，GATA1可与ORMDL3启动子区结合。结论哮喘患者中GATA1表达增加，其可能调控哮喘易感基因ORMDL3启动子活性及ORMDL3表达。

Abstract

Objective To study the expression of the transcription factor GATA1 in bronchial asthma (referred to as asthma) and its effect on the expression level of the asthma susceptibility gene orosomucoid 1-like protein 3 (ORMDL3), along with the underlying molecular mechanisms. Methods The study included 28 cases of moderate asthma, 46 cases of severe asthma, and 12 normal controls from the Gene Expression Omnibus (GEO) database. The mRNA expression levels of GATA1 and ORMDL3 were analyzed among the asthma patients and the normal controls, including their correlation. The pGL-185/58 plasmid was co-transfected with GATA1 gene siRNA (si-GATA1 group) and siRNA negative control (si-control group) into BEAS-2B cells. Bioinformatics methods were used to predict GATA1 binding sites in the promoter region of the ORMDL3 gene. The dual-luciferase reporter gene system was employed to assess the promoter activity of ORMDL3, while real-time quantitative PCR and Western blotting were used to measure the mRNA and protein expression levels of GATA1 and ORMDL3. Chromatin immunoprecipitation (ChIP) assays were conducted to determine whether GATA1 binds to the promoter region of ORMDL3. Results The expression levels of GATA1 and ORMDL3 mRNA were significantly higher in the severe asthma group compared to the normal control group (P<0.001). Positive correlations were observed between GATA1 mRNA and ORMDL3 mRNA expression levels in both the moderate and severe asthma groups (r=0.636 and 0.341, respectively; P<0.05). In BEAS-2B cells, the dual-luciferase reporter assay revealed that ORMDL3 promoter luciferase activity, as well as ORMDL3 mRNA and protein expression levels, were lower in the si-GATA1 group compared to the si-control group (P<0.05). ChIP assay results demonstrated that GATA1 could bind to the promoter region of ORMDL3. Conclusions The expression of GATA1 is increased in asthma patients, which may regulate the promoter activity and expression of the asthma susceptibility gene ORMDL3.

导出引用

陈虎, 李娇娇, 袁越, 金蕊. GATA1在支气管哮喘中的表达及对ORMDL3基因转录调控的影响[J]. 中国当代儿科杂志. 2025, 27(2): 212-218 https://doi.org/10.7499/j.issn.1008-8830.2408143

CHEN Hu, LI Jiao-Jiao, YUAN Yue, JIN Rui. Expression of GATA1 in bronchial asthma and its effect on the transcription regulation of the ORMDL3 gene[J]. Chinese Journal of Contemporary Pediatrics. 2025, 27(2): 212-218 https://doi.org/10.7499/j.issn.1008-8830.2408143

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