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Methotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis
We aimed to investigate the effect of methotrexate (MTX) on microRNA modulation in rheumatoid arthritis fibroblast-like synovial cells (RA-FLS). RA-FLS were treated with MTX for 48 h. We then performed miRNA array analysis to investigate differentially expressed miRNAs. Transfection with miR-877-3p...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8584010/ https://www.ncbi.nlm.nih.gov/pubmed/34768991 http://dx.doi.org/10.3390/ijms222111561 |
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author | Iwamoto, Naoki Furukawa, Kaori Endo, Yushiro Shimizu, Toshimasa Sumiyoshi, Remi Umeda, Masataka Koga, Tomohiro Kawashiri, Shin-ya Igawa, Takashi Ichinose, Kunihiro Tamai, Mami Origuchi, Tomoki Kawakami, Atsushi |
author_facet | Iwamoto, Naoki Furukawa, Kaori Endo, Yushiro Shimizu, Toshimasa Sumiyoshi, Remi Umeda, Masataka Koga, Tomohiro Kawashiri, Shin-ya Igawa, Takashi Ichinose, Kunihiro Tamai, Mami Origuchi, Tomoki Kawakami, Atsushi |
author_sort | Iwamoto, Naoki |
collection | PubMed |
description | We aimed to investigate the effect of methotrexate (MTX) on microRNA modulation in rheumatoid arthritis fibroblast-like synovial cells (RA-FLS). RA-FLS were treated with MTX for 48 h. We then performed miRNA array analysis to investigate differentially expressed miRNAs. Transfection with miR-877-3p precursor and inhibitor were used to investigate the functional role of miR-877-3p in RA-FLS. Gene ontology analysis was used to investigate the cellular processes involving miR-877-3p. The production of cytokines/chemokines was screened by multiplex cytokine/chemokine bead assay and confirmed by ELISA and quantitative real-time PCR. The migratory and proliferative activities of RA-FLS were analyzed by wound healing assay and MKI-67 expression. MTX treatment altered the expression of 13 miRNAs (seven were upregulated and six were downregulated). Among them, quantitative real-time PCR confirmed that miR-877-3p was upregulated in response to MTX (1.79 ± 0.46-fold, p < 0.05). The possible target genes of miR-877-3p in RA-FLS revealed by the microarray analysis were correlated with biological processes. The overexpression of miR-877-3p decreased the production of GM-CSF and CCL3, and the overexpression of miR-877-3p inhibited migratory and proliferative activity. MTX altered the miR-877-3p expression on RA-FLS, and this alteration of miR-877-3p attenuated the abundant production of cytokines/chemokines and proliferative property of RA-FLS. |
format | Online Article Text |
id | pubmed-8584010 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-85840102021-11-12 Methotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis Iwamoto, Naoki Furukawa, Kaori Endo, Yushiro Shimizu, Toshimasa Sumiyoshi, Remi Umeda, Masataka Koga, Tomohiro Kawashiri, Shin-ya Igawa, Takashi Ichinose, Kunihiro Tamai, Mami Origuchi, Tomoki Kawakami, Atsushi Int J Mol Sci Brief Report We aimed to investigate the effect of methotrexate (MTX) on microRNA modulation in rheumatoid arthritis fibroblast-like synovial cells (RA-FLS). RA-FLS were treated with MTX for 48 h. We then performed miRNA array analysis to investigate differentially expressed miRNAs. Transfection with miR-877-3p precursor and inhibitor were used to investigate the functional role of miR-877-3p in RA-FLS. Gene ontology analysis was used to investigate the cellular processes involving miR-877-3p. The production of cytokines/chemokines was screened by multiplex cytokine/chemokine bead assay and confirmed by ELISA and quantitative real-time PCR. The migratory and proliferative activities of RA-FLS were analyzed by wound healing assay and MKI-67 expression. MTX treatment altered the expression of 13 miRNAs (seven were upregulated and six were downregulated). Among them, quantitative real-time PCR confirmed that miR-877-3p was upregulated in response to MTX (1.79 ± 0.46-fold, p < 0.05). The possible target genes of miR-877-3p in RA-FLS revealed by the microarray analysis were correlated with biological processes. The overexpression of miR-877-3p decreased the production of GM-CSF and CCL3, and the overexpression of miR-877-3p inhibited migratory and proliferative activity. MTX altered the miR-877-3p expression on RA-FLS, and this alteration of miR-877-3p attenuated the abundant production of cytokines/chemokines and proliferative property of RA-FLS. MDPI 2021-10-26 /pmc/articles/PMC8584010/ /pubmed/34768991 http://dx.doi.org/10.3390/ijms222111561 Text en © 2021 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 | Brief Report Iwamoto, Naoki Furukawa, Kaori Endo, Yushiro Shimizu, Toshimasa Sumiyoshi, Remi Umeda, Masataka Koga, Tomohiro Kawashiri, Shin-ya Igawa, Takashi Ichinose, Kunihiro Tamai, Mami Origuchi, Tomoki Kawakami, Atsushi Methotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis |
title | Methotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis |
title_full | Methotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis |
title_fullStr | Methotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis |
title_full_unstemmed | Methotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis |
title_short | Methotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis |
title_sort | methotrexate alters the expression of microrna in fibroblast-like synovial cells in rheumatoid arthritis |
topic | Brief Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8584010/ https://www.ncbi.nlm.nih.gov/pubmed/34768991 http://dx.doi.org/10.3390/ijms222111561 |
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