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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
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.
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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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