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miR-23a-3p regulates the proliferation and apoptosis of human lens epithelial cells by targeting Bcl-2 in an in vitro model of cataracts

Cataracts account for ~50% of the cases of blindness in individuals worldwide. The apoptosis of lens epithelial cells (LECs) occurs during the formation of cataracts, which is a non-congenital condition. Numerous microRNAs (miRs) have been reported to regulate apoptosis in LECs. For instance, miR-23...

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Autores principales: Yao, Pengxiang, Jiang, Jian, Ma, Xiaoping, Chen, Zhenzhong, Hong, Yufang, Wu, Yang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7967796/
https://www.ncbi.nlm.nih.gov/pubmed/33777189
http://dx.doi.org/10.3892/etm.2021.9853
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author Yao, Pengxiang
Jiang, Jian
Ma, Xiaoping
Chen, Zhenzhong
Hong, Yufang
Wu, Yang
author_facet Yao, Pengxiang
Jiang, Jian
Ma, Xiaoping
Chen, Zhenzhong
Hong, Yufang
Wu, Yang
author_sort Yao, Pengxiang
collection PubMed
description Cataracts account for ~50% of the cases of blindness in individuals worldwide. The apoptosis of lens epithelial cells (LECs) occurs during the formation of cataracts, which is a non-congenital condition. Numerous microRNAs (miRs) have been reported to regulate apoptosis in LECs. For instance, miR-23a expression levels were shown to be upregulated in cataractous lenses; however, the function of miR-23a in cataracts remains undetermined. To establish an in vitro model of cataracts, human LECs, HLE-B3 cells, were induced with 200 µmol/l H(2)O(2) for 24 h. HLE-B3 cells were transfected with the miR-negative control (NC) mimic, miR-23a-3p mimic, miR-NC inhibitor, miR-23a-3p inhibitor, small interfering RNA (siRNA) targeting BCL2 (siRNA-BCL2) and siRNA-NC. The expression levels of miR-23a-3p were detected using reverse transcription-quantitative PCR. The interaction between miR-23a-3p and the 3'-untranslated region (UTR) of the target mRNA BCL2 was predicted by TargetScan 7.1, and further validated using a dual luciferase reporter assay. The BCL2 protein expression levels were analyzed using western blotting, cell proliferation was determined using a CCK-8 assay and the levels of cell apoptosis were analyzed using flow cytometric analysis. The results of the present study revealed that the expression levels of miR-23a-3p were significantly upregulated, while the expression levels of BCL2 were significantly downregulated in H(2)O(2)-induced HLE-B3 cells compared to untreated control cells. BCL2 was shown to be a target of miR-23a-3p. The miR-23a-3p inhibitor subsequently attenuated H(2)O(2)-induced apoptosis and increased the proliferation of HLE-B3 cells, which was partially reversed by siRNA-BCL2. In conclusion, the findings of the current study suggested that the inhibition of miR-23a-3p may attenuate H(2)O(2)-induced cataract formation by targeting BCL2, thus providing a novel therapeutic target for the treatment of patients with cataracts in the clinic.
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spelling pubmed-79677962021-03-25 miR-23a-3p regulates the proliferation and apoptosis of human lens epithelial cells by targeting Bcl-2 in an in vitro model of cataracts Yao, Pengxiang Jiang, Jian Ma, Xiaoping Chen, Zhenzhong Hong, Yufang Wu, Yang Exp Ther Med Articles Cataracts account for ~50% of the cases of blindness in individuals worldwide. The apoptosis of lens epithelial cells (LECs) occurs during the formation of cataracts, which is a non-congenital condition. Numerous microRNAs (miRs) have been reported to regulate apoptosis in LECs. For instance, miR-23a expression levels were shown to be upregulated in cataractous lenses; however, the function of miR-23a in cataracts remains undetermined. To establish an in vitro model of cataracts, human LECs, HLE-B3 cells, were induced with 200 µmol/l H(2)O(2) for 24 h. HLE-B3 cells were transfected with the miR-negative control (NC) mimic, miR-23a-3p mimic, miR-NC inhibitor, miR-23a-3p inhibitor, small interfering RNA (siRNA) targeting BCL2 (siRNA-BCL2) and siRNA-NC. The expression levels of miR-23a-3p were detected using reverse transcription-quantitative PCR. The interaction between miR-23a-3p and the 3'-untranslated region (UTR) of the target mRNA BCL2 was predicted by TargetScan 7.1, and further validated using a dual luciferase reporter assay. The BCL2 protein expression levels were analyzed using western blotting, cell proliferation was determined using a CCK-8 assay and the levels of cell apoptosis were analyzed using flow cytometric analysis. The results of the present study revealed that the expression levels of miR-23a-3p were significantly upregulated, while the expression levels of BCL2 were significantly downregulated in H(2)O(2)-induced HLE-B3 cells compared to untreated control cells. BCL2 was shown to be a target of miR-23a-3p. The miR-23a-3p inhibitor subsequently attenuated H(2)O(2)-induced apoptosis and increased the proliferation of HLE-B3 cells, which was partially reversed by siRNA-BCL2. In conclusion, the findings of the current study suggested that the inhibition of miR-23a-3p may attenuate H(2)O(2)-induced cataract formation by targeting BCL2, thus providing a novel therapeutic target for the treatment of patients with cataracts in the clinic. D.A. Spandidos 2021-05 2021-02-26 /pmc/articles/PMC7967796/ /pubmed/33777189 http://dx.doi.org/10.3892/etm.2021.9853 Text en Copyright: © Yao et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Yao, Pengxiang
Jiang, Jian
Ma, Xiaoping
Chen, Zhenzhong
Hong, Yufang
Wu, Yang
miR-23a-3p regulates the proliferation and apoptosis of human lens epithelial cells by targeting Bcl-2 in an in vitro model of cataracts
title miR-23a-3p regulates the proliferation and apoptosis of human lens epithelial cells by targeting Bcl-2 in an in vitro model of cataracts
title_full miR-23a-3p regulates the proliferation and apoptosis of human lens epithelial cells by targeting Bcl-2 in an in vitro model of cataracts
title_fullStr miR-23a-3p regulates the proliferation and apoptosis of human lens epithelial cells by targeting Bcl-2 in an in vitro model of cataracts
title_full_unstemmed miR-23a-3p regulates the proliferation and apoptosis of human lens epithelial cells by targeting Bcl-2 in an in vitro model of cataracts
title_short miR-23a-3p regulates the proliferation and apoptosis of human lens epithelial cells by targeting Bcl-2 in an in vitro model of cataracts
title_sort mir-23a-3p regulates the proliferation and apoptosis of human lens epithelial cells by targeting bcl-2 in an in vitro model of cataracts
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7967796/
https://www.ncbi.nlm.nih.gov/pubmed/33777189
http://dx.doi.org/10.3892/etm.2021.9853
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