Functional characterization of rs2229094 (T>C) polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the Retina 4 project

Functional characterization of rs2229094 (T>C) polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the Retina 4 project

Salvador Pastor-Idoate,1,2 Irene Rodríguez-Hernández,2,3 Jimena Rojas,1 Lucia Gonzalez-Buendia,1 Santiago Delgado-Tirado,1,4 Jose Carlos López,1 Rogelio González-Sarmiento,2,3 Jose C Pastor1,4 1IOBA Eye Institute, University of Valladolid, Valladolid, 2Mo...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Pastor-Idoate S, Rodríguez-Hernández I, Rojas J, Gonzalez-Buendia L, Delgado-Tirado S, Lopez JC, González-Sarmiento R, Pastor JC
Formato: article
Lenguaje:EN
Publicado: Dove Medical Press 2017
Materias:
Proliferative vitreoretinopathy
Lymphotoxin alpha
Tumor necrosis factor alpha
inflammation
cytokines
polymorphism.
Ophthalmology
RE1-994
Acceso en línea:https://doaj.org/article/13020835cb2b49308f5a13493aa75269
Etiquetas: Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
id oai:doaj.org-article:13020835cb2b49308f5a13493aa75269
record_format dspace
spelling oai:doaj.org-article:13020835cb2b49308f5a13493aa752692021-12-02T06:09:15ZFunctional characterization of rs2229094 (T>C) polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the Retina 4 project1177-5483https://doaj.org/article/13020835cb2b49308f5a13493aa752692017-05-01T00:00:00Zhttps://www.dovepress.com/functional-characterization-of-rs2229094-tgtc-polymorphism-in-the-tumo-peer-reviewed-article-OPTHhttps://doaj.org/toc/1177-5483Salvador Pastor-Idoate,1,2 Irene Rodríguez-Hernández,2,3 Jimena Rojas,1 Lucia Gonzalez-Buendia,1 Santiago Delgado-Tirado,1,4 Jose Carlos López,1 Rogelio González-Sarmiento,2,3 Jose C Pastor1,4 1IOBA Eye Institute, University of Valladolid, Valladolid, 2Molecular Medicine Unit, Department of Medicine, 3Molecular and Cellular Cancer Biology Institute, High Council of Scientific Research, Biomedical Research Institute of Salamanca, University of Salamanca, Salamanca, 4Department of Ophthalmology, Hospital Clínico Universitario, Valladolid, Spain Purpose: The objective of this study is to determine the expression and localization of lymphotoxin alpha (LTA) in human retinas and the functionality of one of its polymorphisms rs2229094 (C13R) (T>C), previously associated with proliferative vitreoretinopathy (PVR) development.Materials and methods: Total RNA from three healthy human retinas were extracted and subjected to reverse transcription-polymerase chain reaction (RT-PCR) analysis, using flanking primers of LTA cDNA. In addition, three human eyes with retinal detachment (RD) and three healthy control eyes were subjected to immunohistochemistry (IHC) with a specific antibody against LTA. The functionality of T and C alleles was assessed by using pCEFL-Flag expression vector and transient transfection assays in COS-1 cell line. In addition, expression analysis by RT-PCR, Western blot and subcellular localization of both alleles and by immunofluorescence assay was performed.Results: RT-PCR analysis revealed no significant levels of messenger RNA (mRNA) LTA in healthy human retinas. Sequential IHC staining showed differences between healthy human and RD retinas. No differences in mRNA and protein expression levels and in subcellular localization between both alleles were found. Both alleles were located in the cytoplasm of COS-1 cells.Conclusion: Although results suggest lack of functionality, the differences found in IHC study and its strong association with PVR and its relationship with tumor necrosis factor locus, warrant further studies and could justify the use of this polymorphism as a valid biomarker to identify high-risk patients to develop PVR after RD. Keywords: proliferative vitreoretinopathy, lymphotoxin alpha, tumor necrosis factor alpha, inflammation, cytokines, polymorphismPastor-Idoate SRodríguez-Hernández IRojas JGonzalez-Buendia LDelgado-Tirado SLopez JCGonzález-Sarmiento RPastor JCDove Medical PressarticleProliferative vitreoretinopathyLymphotoxin alphaTumor necrosis factor alphainflammationcytokinespolymorphism.OphthalmologyRE1-994ENClinical Ophthalmology, Vol Volume 11, Pp 973-981 (2017)
institution DOAJ
collection DOAJ
language EN
topic Proliferative vitreoretinopathy
Lymphotoxin alpha
Tumor necrosis factor alpha
inflammation
cytokines
polymorphism.
Ophthalmology
RE1-994
spellingShingle Proliferative vitreoretinopathy
Lymphotoxin alpha
Tumor necrosis factor alpha
inflammation
cytokines
polymorphism.
Ophthalmology
RE1-994
Pastor-Idoate S
Rodríguez-Hernández I
Rojas J
Gonzalez-Buendia L
Delgado-Tirado S
Lopez JC
González-Sarmiento R
Pastor JC
Functional characterization of rs2229094 (T>C) polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the Retina 4 project
description Salvador Pastor-Idoate,1,2 Irene Rodríguez-Hernández,2,3 Jimena Rojas,1 Lucia Gonzalez-Buendia,1 Santiago Delgado-Tirado,1,4 Jose Carlos López,1 Rogelio González-Sarmiento,2,3 Jose C Pastor1,4 1IOBA Eye Institute, University of Valladolid, Valladolid, 2Molecular Medicine Unit, Department of Medicine, 3Molecular and Cellular Cancer Biology Institute, High Council of Scientific Research, Biomedical Research Institute of Salamanca, University of Salamanca, Salamanca, 4Department of Ophthalmology, Hospital Clínico Universitario, Valladolid, Spain Purpose: The objective of this study is to determine the expression and localization of lymphotoxin alpha (LTA) in human retinas and the functionality of one of its polymorphisms rs2229094 (C13R) (T>C), previously associated with proliferative vitreoretinopathy (PVR) development.Materials and methods: Total RNA from three healthy human retinas were extracted and subjected to reverse transcription-polymerase chain reaction (RT-PCR) analysis, using flanking primers of LTA cDNA. In addition, three human eyes with retinal detachment (RD) and three healthy control eyes were subjected to immunohistochemistry (IHC) with a specific antibody against LTA. The functionality of T and C alleles was assessed by using pCEFL-Flag expression vector and transient transfection assays in COS-1 cell line. In addition, expression analysis by RT-PCR, Western blot and subcellular localization of both alleles and by immunofluorescence assay was performed.Results: RT-PCR analysis revealed no significant levels of messenger RNA (mRNA) LTA in healthy human retinas. Sequential IHC staining showed differences between healthy human and RD retinas. No differences in mRNA and protein expression levels and in subcellular localization between both alleles were found. Both alleles were located in the cytoplasm of COS-1 cells.Conclusion: Although results suggest lack of functionality, the differences found in IHC study and its strong association with PVR and its relationship with tumor necrosis factor locus, warrant further studies and could justify the use of this polymorphism as a valid biomarker to identify high-risk patients to develop PVR after RD. Keywords: proliferative vitreoretinopathy, lymphotoxin alpha, tumor necrosis factor alpha, inflammation, cytokines, polymorphism
format article
author Pastor-Idoate S
Rodríguez-Hernández I
Rojas J
Gonzalez-Buendia L
Delgado-Tirado S
Lopez JC
González-Sarmiento R
Pastor JC
author_facet Pastor-Idoate S
Rodríguez-Hernández I
Rojas J
Gonzalez-Buendia L
Delgado-Tirado S
Lopez JC
González-Sarmiento R
Pastor JC
author_sort Pastor-Idoate S
title Functional characterization of rs2229094 (T>C) polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the Retina 4 project
title_short Functional characterization of rs2229094 (T>C) polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the Retina 4 project
title_full Functional characterization of rs2229094 (T>C) polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the Retina 4 project
title_fullStr Functional characterization of rs2229094 (T>C) polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the Retina 4 project
title_full_unstemmed Functional characterization of rs2229094 (T>C) polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the Retina 4 project
title_sort functional characterization of rs2229094 (t>c) polymorphism in the tumor necrosis factor locus and lymphotoxin alpha expression in human retina: the retina 4 project
publisher Dove Medical Press
publishDate 2017
url https://doaj.org/article/13020835cb2b49308f5a13493aa75269
work_keys_str_mv AT pastoridoates functionalcharacterizationofrs2229094tgtcpolymorphisminthetumornecrosisfactorlocusandlymphotoxinalphaexpressioninhumanretinatheretina4project
AT rodriguezhernandezi functionalcharacterizationofrs2229094tgtcpolymorphisminthetumornecrosisfactorlocusandlymphotoxinalphaexpressioninhumanretinatheretina4project
AT rojasj functionalcharacterizationofrs2229094tgtcpolymorphisminthetumornecrosisfactorlocusandlymphotoxinalphaexpressioninhumanretinatheretina4project
AT gonzalezbuendial functionalcharacterizationofrs2229094tgtcpolymorphisminthetumornecrosisfactorlocusandlymphotoxinalphaexpressioninhumanretinatheretina4project
AT delgadotirados functionalcharacterizationofrs2229094tgtcpolymorphisminthetumornecrosisfactorlocusandlymphotoxinalphaexpressioninhumanretinatheretina4project
AT lopezjc functionalcharacterizationofrs2229094tgtcpolymorphisminthetumornecrosisfactorlocusandlymphotoxinalphaexpressioninhumanretinatheretina4project
AT gonzalezsarmientor functionalcharacterizationofrs2229094tgtcpolymorphisminthetumornecrosisfactorlocusandlymphotoxinalphaexpressioninhumanretinatheretina4project
AT pastorjc functionalcharacterizationofrs2229094tgtcpolymorphisminthetumornecrosisfactorlocusandlymphotoxinalphaexpressioninhumanretinatheretina4project
_version_ 1718400035990798336

Ejemplares similares