Dragicevic Sandra, Milosevic Katarina, Nestorovic Branimir, Nikolic Aleksandra
Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia.
Department of Pulmonology and Allergology, University Children's Hospital, Belgrade, Serbia.
Pediatr Allergy Immunol Pulmonol. 2017 Dec;30(4):239-245. doi: 10.1089/ped.2017.0770. Epub 2017 Oct 12.
Transforming growth factor beta 1 (TGFB1) is a multifunctional cytokine with a key role in asthma airway inflammation and remodeling. Since elevated levels of this cytokine in airways might be associated with response to asthma therapy, the aim of this study was to investigate whether the presence of the polymorphism C-509T in the promoter of the gene is associated with response to montelukast. A group of 102 asthmatic patients was genotyped for the presence of the C-509T polymorphism by DNA sequencing and subjected to induced sputum sampling. Cells from sputum samples and BEAS 2B cells were treated with montelukast and endogenous expression was measured by quantitative real-time polymerase chain reaction. The promoter activity was analyzed by luciferase assays in BEAS 2B cells transfected with constructs carrying variants -509C and -509T of the gene promoter. After treatment with montelukast, the decrease in gene expression was greater for the -509TT genotype (58.9%) than for the -509CC and -509CT genotypes (49.6% and 31.8%, respectively) ( = 0.071). In BEAS 2B cells, expression of endogenous was reduced by about 27% after montelukast treatment, while luciferase activity of both promoter variants was increased after montelukast treatment (-509C allele: 48.3%, = 0.060; and -509T allele: 100.5%, = 0.062). A more intensive response was registered in the promoter containing the -509T allele, which had 135% higher activity than the -509C variant ( = 0.035). This study showed that the presence of the -509T allele in the promoter might modulate effects of montelukast on gene expression, but future studies are necessary, taking into consideration other genetic and nongenetic factors. It is of potential importance for clinical management of asthma to clarify the influence of the C-509T polymorphism on the response to treatment with montelukast.
转化生长因子β1（TGFB1）是一种多功能细胞因子，在哮喘气道炎症和重塑中起关键作用。由于气道中这种细胞因子水平升高可能与哮喘治疗反应有关，本研究的目的是调查该基因启动子中C-509T多态性的存在是否与孟鲁司特的反应有关。通过DNA测序对一组102例哮喘患者进行C-509T多态性基因分型，并进行诱导痰采样。用孟鲁司特处理痰样本细胞和BEAS 2B细胞，并通过定量实时聚合酶链反应测量内源性表达。通过荧光素酶测定分析用携带该基因启动子变体-509C和-509T的构建体转染的BEAS 2B细胞中的启动子活性。用孟鲁司特处理后，-509TT基因型的基因表达下降幅度（58.9%）大于-509CC和-509CT基因型（分别为49.6%和31.8%）（P = 0.071）。在BEAS 2B细胞中，孟鲁司特处理后内源性表达降低约27%，而两种启动子变体的荧光素酶活性在孟鲁司特处理后均增加（-509C等位基因：48.3%，P = 0.060；-509T等位基因：100.5%，P = 0.062）。在含有-509T等位基因的启动子中观察到更强的反应，其活性比-509C变体高135%（P = 0.035）。本研究表明，该基因启动子中-509T等位基因的存在可能调节孟鲁司特对该基因表达的影响，但考虑到其他遗传和非遗传因素，未来仍需进行研究。阐明C-509T多态性对孟鲁司特治疗反应的影响对哮喘的临床管理具有潜在重要性。
