PTGDR gene expression and response to dexamethasone treatment in an in vitro model

• DOI: 10.1371/journal.pone.0186957
• Publication Date: 2017-10-31T14:17:09Z
• Authors: Elena Marcos-Vadillo; Asunción García-S...; Catalina Sanz; Ignacio Davila; María Isidoro-García
• Keywords: 0301 basic medicine; Lung Neoplasms; Science; Receptors, Prostaglandin; Adenocarcinoma; Dexamethasone; 03 medical and health sciences; Humans; Receptors, Immunologic; Promoter Regions, Genetic; Glucocorticoids; Asma; 0303 health sciences; Base Sequence; Reverse Transcriptase Polymerase Chain Reaction; Q; Expresi?n g?nica; R; Epithelial Cells; Flow Cytometry; Asthma; 3. Good health; Gene Expression Regulation, Neoplastic; Haplotypes; A549 Cells; Medicine; Cytokines; Gene expression; Expresión génica; Research Article

Asthma is a multifactorial pathology influenced by environmental and genetic factors. Glucocorticoid treatment decreases symptoms by regulating genes involved in the inflammatory process through binding to specific DNA sequences. Polymorphisms located in the promoter region of the Prostaglandin D Receptor (PTGDR) gene have been related to asthma. We aimed to analyze the effect of PTGDR promoter haplotypes on gene expression and response to corticosteroid therapy. A549 lung epithelial cells were transfected with vectors carrying four different PTGDR haplotypes (CTCT, CCCC, CCCT and TCCT), and treated with dexamethasone. Different approaches to study the promoter activity (Dual Luciferase Reporter System), gene expression levels (qPCR) and cytokine secretion (Multiplexed Bead-based Flow Cytometric) were used. In addition, in silico analysis was also performed. Cells carrying the TCCT haplotype showed the lowest promoter activity (p-value<0.05) and mRNA expression levels in basal conditions. After dexamethasone treatment, cells carrying the wild-type variant CTCT showed the highest response, and those carrying the TCCT variant the lowest (p-value<0.05) in luciferase assays. Different transcription factor binding patterns were identified in silico. Moreover, differences in cytokine secretion were also found among different promoter haplotypes. Polymorphisms of PTGDR gene influence basal promoter activity and gene expression, as well as the cytokine secretory pattern. Furthermore, an association between these positions and response to corticoid treatment was observed.