Retinoic acid selectively modulated PTGDR expression in A549 lungcarcinoma cell line
- García-Sánchez, A 1
- Marcos-Vadillo, E 2
- Hernández-Hernández, L 2
- Sanz, C 3
- Laffond, E 12
- Gracia, T 2
- Lorente, F 12
- Dávila, I 12
- Isidoro-García, M 12
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1
Universidad de Salamanca
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2
Hospital Universitario de Salamanca
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- 3 Departamento de Microbiología, Universidad de Salamanca
ISSN: 0105-4538, 1398-9995
Year of publication: 2015
Volume: 70
Issue: s101
Pages: 23-23
Congress: European Academy of Allergy and Clinical Immunology Congress
Type: Conference paper
Abstract
Background:Asthma and allergy are com-plex diseases that results from interactionbetween genetic background and environ-mental factors. Retinoic Acid is a vitaminA derivative which has been involved inthe development of immunological diseasesalthough its role has not been established.RA has previously been associated to pros-taglandin pathway.PTGDR, a receptor ofPGD2 has been reported as a candidategene in asthma. The goal of this study isto determine a putative role of RA inasthma development throughPTGDRacti-vation, to innovate and improve diagnosticand therapeutic strategies.Method:A549 cell line was transfectedwith four pGL3-PTGDRreporter con-tructs. Seven hundred pb from homozy-gous subjects who had the followingalternative haplotypes (positions 613, 549, 441 and 197): CTCT, CCCT,TCCT and CCCC), were cloned in intofirefly luciferase pGL3-Basic vector. Aftertransfection cells were treated with0.01lM and 1lM all-trans-Retinoic Acid(ATRA) and measured at 24 and 48 hafter treatment. ThePTGDRpromoteractivity was analyzed using the Dual Luci-ferase Reporter System. Firefly luciferasemeasurements were normalized to Renillaluciferase measurements as a control fortransfection efficiency.Results:We examined the transcriptionalactivity ofPTGDRto determine whetherATRA increasesPTGDRpromoter activityin the genetic variants. When stimulatedwith 1lM ATRA at 48 h, CCCC-PTGDRluc exhibited the highest luciferaseactivity (2.06 0.53). A nearly similarincrease in the luciferase activity was alsoobserved in CCCT-PTGDRluc and TCCT-PTGDRluc (1.98 0.32; and 1.97 0.27).This increase ofPTGDRactivation byATRA stimulation was detected in thevariants when the 549C mutated variantwas present. No increase was observed forthe wild type variant CTCT (613C, 549T, 441C, and 197T) (P<0.0001).Conclusion:Ours results indicate that theeffect of Retinoic Acid overPTGDR gene expression selectively depend on the geneticvariant present in its promoter region. Thisfact that could help to explain the contro-versial results reported so far regarding dif-ferential response to RA.