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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2021
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oai:doaj.org-article:a033ad1617144b1a895f980fda3463912021-11-11T17:02:38ZMethotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis10.3390/ijms2221115611422-00671661-6596https://doaj.org/article/a033ad1617144b1a895f980fda3463912021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11561https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067We 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, <i>p</i> < 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.Naoki IwamotoKaori FurukawaYushiro EndoToshimasa ShimizuRemi SumiyoshiMasataka UmedaTomohiro KogaShin-ya KawashiriTakashi IgawaKunihiro IchinoseMami TamaiTomoki OriguchiAtsushi KawakamiMDPI AGarticleMTXmiR-877-3prheumatoid arthritisfibroblast-like synovial cellsmicroRNABiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11561, p 11561 (2021) |
| institution |
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| collection |
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| language |
EN |
| topic |
MTX miR-877-3p rheumatoid arthritis fibroblast-like synovial cells microRNA Biology (General) QH301-705.5 Chemistry QD1-999 |
| spellingShingle |
MTX miR-877-3p rheumatoid arthritis fibroblast-like synovial cells microRNA Biology (General) QH301-705.5 Chemistry QD1-999 Naoki Iwamoto Kaori Furukawa Yushiro Endo Toshimasa Shimizu Remi Sumiyoshi Masataka Umeda Tomohiro Koga Shin-ya Kawashiri Takashi Igawa Kunihiro Ichinose Mami Tamai Tomoki Origuchi Atsushi Kawakami Methotrexate Alters the Expression of microRNA in Fibroblast-like Synovial Cells in Rheumatoid Arthritis |
| 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, <i>p</i> < 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 |
article |
| author |
Naoki Iwamoto Kaori Furukawa Yushiro Endo Toshimasa Shimizu Remi Sumiyoshi Masataka Umeda Tomohiro Koga Shin-ya Kawashiri Takashi Igawa Kunihiro Ichinose Mami Tamai Tomoki Origuchi Atsushi Kawakami |
| author_facet |
Naoki Iwamoto Kaori Furukawa Yushiro Endo Toshimasa Shimizu Remi Sumiyoshi Masataka Umeda Tomohiro Koga Shin-ya Kawashiri Takashi Igawa Kunihiro Ichinose Mami Tamai Tomoki Origuchi Atsushi Kawakami |
| author_sort |
Naoki Iwamoto |
| title |
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_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_sort |
methotrexate alters the expression of microrna in fibroblast-like synovial cells in rheumatoid arthritis |
| publisher |
MDPI AG |
| publishDate |
2021 |
| url |
https://doaj.org/article/a033ad1617144b1a895f980fda346391 |
| work_keys_str_mv |
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1718432192477003776 |