MiR-494-3p induced by compressive force inhibits cell proliferation in MC3T3-E1 cells

Mechanical stimuli regulate fundamental cell processes such as proliferation, differentiation, and morphogenesis. We attempted to identify microRNA (miRNA) whose expression is changed during compressive treatment in MC3T3-E1, a pre-osteoblastic cell line. Microarray analysis followed by reverse tran...

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Veröffentlicht in:Journal of bioscience and bioengineering 2015-10, Vol.120 (4), p.456-462
Hauptverfasser: Iwawaki, Yuki, Mizusawa, Noriko, Iwata, Takeo, Higaki, Nobuaki, Goto, Takaharu, Watanabe, Megumi, Tomotake, Yoritoki, Ichikawa, Tetsuo, Yoshimoto, Katsuhiko
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container_issue 4
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container_title Journal of bioscience and bioengineering
container_volume 120
creator Iwawaki, Yuki
Mizusawa, Noriko
Iwata, Takeo
Higaki, Nobuaki
Goto, Takaharu
Watanabe, Megumi
Tomotake, Yoritoki
Ichikawa, Tetsuo
Yoshimoto, Katsuhiko
description Mechanical stimuli regulate fundamental cell processes such as proliferation, differentiation, and morphogenesis. We attempted to identify microRNA (miRNA) whose expression is changed during compressive treatment in MC3T3-E1, a pre-osteoblastic cell line. Microarray analysis followed by reverse transcription-quantitative polymerase chain reaction revealed that compressive force at 294 Pa for 24 h in MC3T3-E1 cells increased levels of miR-494-3p, miR-146a-5p, miR-210-3p, and miR-1247-3p. Among these miRNAs, miR-494-3p was found to inhibit cell proliferation in MC3T3-E1 cells. Furthermore, cells subjected to compressive force showed slower cell growth compared with control cells. Levels of mRNA for fibroblast growth factor receptor 2 (FGFR2) and Rho-associated coiled-coil kinase 1 (ROCK1), which were predicted to be targets of miR-494-3p, were decreased by compressive force or overexpression of miR-494-3p mimics in MC3T3-E1 cells. Furthermore, binding sites of miR-494-3p within 3′-untranslated regions of Fgfr2 and Rock1 were determined using luciferase reporter assay. In conclusion, compressive force affected expressions of several miRNAs including miR-494-3p in MC3T3-E1 cells. Compressive force might inhibit cell proliferation in osteoblasts by up-regulating miR-494-3p followed by FGFR2 and ROCK1 gene repressions. •Compressive force induced expression of miR-146a-5p, miR-210-3p, miR-494-3p, and miR-1247-3p in MC3T3-E1 cells.•MiR-494-3p inhibited cell proliferation in MC3T3-E1 cells.•FGFR2 and ROCK1 were targets of miR-494-3p in MC3T3-E1 cells.
doi_str_mv 10.1016/j.jbiosc.2015.02.006
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We attempted to identify microRNA (miRNA) whose expression is changed during compressive treatment in MC3T3-E1, a pre-osteoblastic cell line. Microarray analysis followed by reverse transcription-quantitative polymerase chain reaction revealed that compressive force at 294 Pa for 24 h in MC3T3-E1 cells increased levels of miR-494-3p, miR-146a-5p, miR-210-3p, and miR-1247-3p. Among these miRNAs, miR-494-3p was found to inhibit cell proliferation in MC3T3-E1 cells. Furthermore, cells subjected to compressive force showed slower cell growth compared with control cells. Levels of mRNA for fibroblast growth factor receptor 2 (FGFR2) and Rho-associated coiled-coil kinase 1 (ROCK1), which were predicted to be targets of miR-494-3p, were decreased by compressive force or overexpression of miR-494-3p mimics in MC3T3-E1 cells. Furthermore, binding sites of miR-494-3p within 3′-untranslated regions of Fgfr2 and Rock1 were determined using luciferase reporter assay. 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Compressive force might inhibit cell proliferation in osteoblasts by up-regulating miR-494-3p followed by FGFR2 and ROCK1 gene repressions. •Compressive force induced expression of miR-146a-5p, miR-210-3p, miR-494-3p, and miR-1247-3p in MC3T3-E1 cells.•MiR-494-3p inhibited cell proliferation in MC3T3-E1 cells.•FGFR2 and ROCK1 were targets of miR-494-3p in MC3T3-E1 cells.</description><identifier>ISSN: 1389-1723</identifier><identifier>EISSN: 1347-4421</identifier><identifier>DOI: 10.1016/j.jbiosc.2015.02.006</identifier><identifier>PMID: 25795570</identifier><language>eng</language><publisher>Japan: Elsevier B.V</publisher><subject>3' Untranslated Regions - genetics ; Cell Line ; Cell proliferation ; Cell Proliferation - genetics ; Compressive force ; Down-Regulation ; Gene Expression Regulation ; Humans ; Luciferases - genetics ; Mechanical stimuli ; MicroRNA ; MicroRNAs - biosynthesis ; MicroRNAs - genetics ; Oligonucleotide Array Sequence Analysis ; Osteoblasts ; Osteoblasts - cytology ; Osteoblasts - metabolism ; Receptor, Fibroblast Growth Factor, Type 2 - genetics ; Reverse Transcriptase Polymerase Chain Reaction ; rho-Associated Kinases - genetics ; RNA, Messenger - analysis ; RNA, Messenger - genetics ; Stress, Mechanical ; Up-Regulation</subject><ispartof>Journal of bioscience and bioengineering, 2015-10, Vol.120 (4), p.456-462</ispartof><rights>2015 The Society for Biotechnology, Japan</rights><rights>Copyright © 2015 The Society for Biotechnology, Japan. 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We attempted to identify microRNA (miRNA) whose expression is changed during compressive treatment in MC3T3-E1, a pre-osteoblastic cell line. Microarray analysis followed by reverse transcription-quantitative polymerase chain reaction revealed that compressive force at 294 Pa for 24 h in MC3T3-E1 cells increased levels of miR-494-3p, miR-146a-5p, miR-210-3p, and miR-1247-3p. Among these miRNAs, miR-494-3p was found to inhibit cell proliferation in MC3T3-E1 cells. Furthermore, cells subjected to compressive force showed slower cell growth compared with control cells. Levels of mRNA for fibroblast growth factor receptor 2 (FGFR2) and Rho-associated coiled-coil kinase 1 (ROCK1), which were predicted to be targets of miR-494-3p, were decreased by compressive force or overexpression of miR-494-3p mimics in MC3T3-E1 cells. Furthermore, binding sites of miR-494-3p within 3′-untranslated regions of Fgfr2 and Rock1 were determined using luciferase reporter assay. In conclusion, compressive force affected expressions of several miRNAs including miR-494-3p in MC3T3-E1 cells. Compressive force might inhibit cell proliferation in osteoblasts by up-regulating miR-494-3p followed by FGFR2 and ROCK1 gene repressions. •Compressive force induced expression of miR-146a-5p, miR-210-3p, miR-494-3p, and miR-1247-3p in MC3T3-E1 cells.•MiR-494-3p inhibited cell proliferation in MC3T3-E1 cells.•FGFR2 and ROCK1 were targets of miR-494-3p in MC3T3-E1 cells.</abstract><cop>Japan</cop><pub>Elsevier B.V</pub><pmid>25795570</pmid><doi>10.1016/j.jbiosc.2015.02.006</doi><tpages>7</tpages></addata></record>
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subjects 3' Untranslated Regions - genetics
Cell Line
Cell proliferation
Cell Proliferation - genetics
Compressive force
Down-Regulation
Gene Expression Regulation
Humans
Luciferases - genetics
Mechanical stimuli
MicroRNA
MicroRNAs - biosynthesis
MicroRNAs - genetics
Oligonucleotide Array Sequence Analysis
Osteoblasts
Osteoblasts - cytology
Osteoblasts - metabolism
Receptor, Fibroblast Growth Factor, Type 2 - genetics
Reverse Transcriptase Polymerase Chain Reaction
rho-Associated Kinases - genetics
RNA, Messenger - analysis
RNA, Messenger - genetics
Stress, Mechanical
Up-Regulation
title MiR-494-3p induced by compressive force inhibits cell proliferation in MC3T3-E1 cells
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