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Endogenous Galectin-1 Modulates Cell Biological Properties of Immortalized Retinal Pigment Epithelial Cells In Vitro

In the eye, an increase in galectin-1 is associated with various chorioretinal diseases, in which retinal pigment epithelium (RPE) cells play a crucial role in disease development and progression. Since little is known about the function of endogenous galectin-1 in these cells, we developed a galect...

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Autores principales: Liesenhoff, Caspar, Paulus, Simon Martin, Havertz, Caroline, Geerlof, Arie, Priglinger, Siegfried, Priglinger, Claudia Sybille, Ohlmann, Andreas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10454680/
https://www.ncbi.nlm.nih.gov/pubmed/37628816
http://dx.doi.org/10.3390/ijms241612635
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author Liesenhoff, Caspar
Paulus, Simon Martin
Havertz, Caroline
Geerlof, Arie
Priglinger, Siegfried
Priglinger, Claudia Sybille
Ohlmann, Andreas
author_facet Liesenhoff, Caspar
Paulus, Simon Martin
Havertz, Caroline
Geerlof, Arie
Priglinger, Siegfried
Priglinger, Claudia Sybille
Ohlmann, Andreas
author_sort Liesenhoff, Caspar
collection PubMed
description In the eye, an increase in galectin-1 is associated with various chorioretinal diseases, in which retinal pigment epithelium (RPE) cells play a crucial role in disease development and progression. Since little is known about the function of endogenous galectin-1 in these cells, we developed a galectin-1-deficient immortalized RPE cell line (ARPE-19-LGALS1(−/−)) using a sgRNA/Cas9 all-in-one expression vector and investigated its cell biological properties. Galectin-1 deficiency was confirmed by Western blot analysis and immunocytochemistry. Cell viability and proliferation were significantly decreased in ARPE-19-LGALS1(−/−) cells when compared to wild-type controls. Further on, an increased attachment of galectin-1-deficient RPE cells was observed by cell adhesion assay when compared to control cells. The diminished viability and proliferation, as well as the enhanced adhesion of galectin-1-deficient ARPE-19 cells, could be blocked, at least in part, by the additional treatment with human recombinant galectin-1. In addition, a significantly reduced migration was detected in ARPE-19-LGALS1(−/−) cells. In comparison to control cells, galectin-1-deficient RPE cells had enhanced expression of sm-α-actin and N-cadherin, whereas expression of E-cadherin showed no significant alteration. Finally, a compensatory expression of galectin-8 mRNA was observed in ARPE-19-LGALS1(−/−) cells. In conclusion, in RPE cells, endogenous galectin-1 has crucial functions for various cell biological processes, including viability, proliferation, migration, adherence, and retaining the epithelial phenotype.
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spelling pubmed-104546802023-08-26 Endogenous Galectin-1 Modulates Cell Biological Properties of Immortalized Retinal Pigment Epithelial Cells In Vitro Liesenhoff, Caspar Paulus, Simon Martin Havertz, Caroline Geerlof, Arie Priglinger, Siegfried Priglinger, Claudia Sybille Ohlmann, Andreas Int J Mol Sci Article In the eye, an increase in galectin-1 is associated with various chorioretinal diseases, in which retinal pigment epithelium (RPE) cells play a crucial role in disease development and progression. Since little is known about the function of endogenous galectin-1 in these cells, we developed a galectin-1-deficient immortalized RPE cell line (ARPE-19-LGALS1(−/−)) using a sgRNA/Cas9 all-in-one expression vector and investigated its cell biological properties. Galectin-1 deficiency was confirmed by Western blot analysis and immunocytochemistry. Cell viability and proliferation were significantly decreased in ARPE-19-LGALS1(−/−) cells when compared to wild-type controls. Further on, an increased attachment of galectin-1-deficient RPE cells was observed by cell adhesion assay when compared to control cells. The diminished viability and proliferation, as well as the enhanced adhesion of galectin-1-deficient ARPE-19 cells, could be blocked, at least in part, by the additional treatment with human recombinant galectin-1. In addition, a significantly reduced migration was detected in ARPE-19-LGALS1(−/−) cells. In comparison to control cells, galectin-1-deficient RPE cells had enhanced expression of sm-α-actin and N-cadherin, whereas expression of E-cadherin showed no significant alteration. Finally, a compensatory expression of galectin-8 mRNA was observed in ARPE-19-LGALS1(−/−) cells. In conclusion, in RPE cells, endogenous galectin-1 has crucial functions for various cell biological processes, including viability, proliferation, migration, adherence, and retaining the epithelial phenotype. MDPI 2023-08-10 /pmc/articles/PMC10454680/ /pubmed/37628816 http://dx.doi.org/10.3390/ijms241612635 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Liesenhoff, Caspar
Paulus, Simon Martin
Havertz, Caroline
Geerlof, Arie
Priglinger, Siegfried
Priglinger, Claudia Sybille
Ohlmann, Andreas
Endogenous Galectin-1 Modulates Cell Biological Properties of Immortalized Retinal Pigment Epithelial Cells In Vitro
title Endogenous Galectin-1 Modulates Cell Biological Properties of Immortalized Retinal Pigment Epithelial Cells In Vitro
title_full Endogenous Galectin-1 Modulates Cell Biological Properties of Immortalized Retinal Pigment Epithelial Cells In Vitro
title_fullStr Endogenous Galectin-1 Modulates Cell Biological Properties of Immortalized Retinal Pigment Epithelial Cells In Vitro
title_full_unstemmed Endogenous Galectin-1 Modulates Cell Biological Properties of Immortalized Retinal Pigment Epithelial Cells In Vitro
title_short Endogenous Galectin-1 Modulates Cell Biological Properties of Immortalized Retinal Pigment Epithelial Cells In Vitro
title_sort endogenous galectin-1 modulates cell biological properties of immortalized retinal pigment epithelial cells in vitro
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10454680/
https://www.ncbi.nlm.nih.gov/pubmed/37628816
http://dx.doi.org/10.3390/ijms241612635
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