期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 24, 期 16, 页码 -出版社
MDPI
DOI: 10.3390/ijms241612635
关键词
galectin-1; galectin-1 deficiency; galectin-1 knockout; retinal pigment epithelium cells; ARPE-19; cell proliferation; cell attachment; cell viability; epithelial-mesenchymal transition; compensatory gene expression
Galectin-1 deficiency in retinal pigment epithelium (RPE) cells affects various cell biological processes, including cell survival, proliferation, migration, adhesion, and maintenance of the epithelial phenotype.
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-a-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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