Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells

OBJECTIVES: This study aimed to investigate the functional effects of microRNA (miR)-214-5p on osteoblastic cells, which might provide a potential role of miR-214-5p in bone fracture healing. METHODS: Blood samples were obtained from patients with hand fracture or intra-articular calcaneal fracture...

Descripción completa

Detalles Bibliográficos
Autores principales: Li, Q. S., Meng, F. Y., Zhao, Y. H., Jin, C. L., Tian, J., Yi, X. J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2017
Materias:
Research
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5579316/
https://www.ncbi.nlm.nih.gov/pubmed/28784704
http://dx.doi.org/10.1302/2046-3758.68.BJR-2016-0208.R2
_version_ 1783260686141358080
author Li, Q. S.
Meng, F. Y.
Zhao, Y. H.
Jin, C. L.
Tian, J.
Yi, X. J.
author_facet Li, Q. S.
Meng, F. Y.
Zhao, Y. H.
Jin, C. L.
Tian, J.
Yi, X. J.
author_sort Li, Q. S.
collection PubMed
description OBJECTIVES: This study aimed to investigate the functional effects of microRNA (miR)-214-5p on osteoblastic cells, which might provide a potential role of miR-214-5p in bone fracture healing. METHODS: Blood samples were obtained from patients with hand fracture or intra-articular calcaneal fracture and from healthy controls (HCs). Expression of miR-214-5p was monitored by qRT-PCR at day 7, 14 and 21 post-surgery. Mouse osteoblastic MC3T3-E1 cells were transfected with antisense oligonucleotides (ASO)-miR-214-5p, collagen type IV alpha 1 (COL4A1) vector or their controls; thereafter, cell viability, apoptotic rate, and the expression of collagen type I alpha 1 (COL1A1), type II collagen (COL-II), and type X collagen (COL-X) were determined. Luciferase reporter assay, qRT-PCR, and Western blot were performed to ascertain whether COL4A1 was a target of miR-214-5p. RESULTS: Plasma miR-214-5p was highly expressed in patients with bone fracture compared with HCs after fracture (p < 0.05 or p < 0.01). Inhibition of miR-214-5p increased the viability of MC3T3-E1 cells and the expressions of COL1A1 and COL-X, but decreased the apoptotic rate and COL-II expression (p < 0.05 or p < 0.01). COL4A1 was a target of miR-214-5p, and was negatively regulated by miR-214-5p (p < 0.05 or p < 0.01). Overexpression of COL4A1 showed a similar impact on cell viability, apoptotic rate, and COL1A1, COL-II, and COL-X expressions inhibiting miR-214-5p (p < 0.01). CONCLUSION: Inhibition of miR-214-5p promotes cell survival and extracellular matrix (ECM) formation of osteoblastic MC3T3-E1 cells by targeting COL4A1. Cite this article: Q. S. Li, F. Y. Meng, Y. H. Zhao, C. L. Jin, J. Tian, X. J. Yi. Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells. Bone Joint Res 2017;6:464–471. DOI: 10.1302/2046-3758.68.BJR-2016-0208.R2
format Online
Article
Text
id pubmed-5579316
institution National Center for Biotechnology Information
language English
publishDate 2017
record_format MEDLINE/PubMed
spelling pubmed-55793162017-09-07 Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells Li, Q. S. Meng, F. Y. Zhao, Y. H. Jin, C. L. Tian, J. Yi, X. J. Bone Joint Res Research OBJECTIVES: This study aimed to investigate the functional effects of microRNA (miR)-214-5p on osteoblastic cells, which might provide a potential role of miR-214-5p in bone fracture healing. METHODS: Blood samples were obtained from patients with hand fracture or intra-articular calcaneal fracture and from healthy controls (HCs). Expression of miR-214-5p was monitored by qRT-PCR at day 7, 14 and 21 post-surgery. Mouse osteoblastic MC3T3-E1 cells were transfected with antisense oligonucleotides (ASO)-miR-214-5p, collagen type IV alpha 1 (COL4A1) vector or their controls; thereafter, cell viability, apoptotic rate, and the expression of collagen type I alpha 1 (COL1A1), type II collagen (COL-II), and type X collagen (COL-X) were determined. Luciferase reporter assay, qRT-PCR, and Western blot were performed to ascertain whether COL4A1 was a target of miR-214-5p. RESULTS: Plasma miR-214-5p was highly expressed in patients with bone fracture compared with HCs after fracture (p < 0.05 or p < 0.01). Inhibition of miR-214-5p increased the viability of MC3T3-E1 cells and the expressions of COL1A1 and COL-X, but decreased the apoptotic rate and COL-II expression (p < 0.05 or p < 0.01). COL4A1 was a target of miR-214-5p, and was negatively regulated by miR-214-5p (p < 0.05 or p < 0.01). Overexpression of COL4A1 showed a similar impact on cell viability, apoptotic rate, and COL1A1, COL-II, and COL-X expressions inhibiting miR-214-5p (p < 0.01). CONCLUSION: Inhibition of miR-214-5p promotes cell survival and extracellular matrix (ECM) formation of osteoblastic MC3T3-E1 cells by targeting COL4A1. Cite this article: Q. S. Li, F. Y. Meng, Y. H. Zhao, C. L. Jin, J. Tian, X. J. Yi. Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells. Bone Joint Res 2017;6:464–471. DOI: 10.1302/2046-3758.68.BJR-2016-0208.R2 2017-09-01 /pmc/articles/PMC5579316/ /pubmed/28784704 http://dx.doi.org/10.1302/2046-3758.68.BJR-2016-0208.R2 Text en © 2017 Yi et al. This is an open-access article distributed under the terms of the Creative Commons Attributions licence (CC-BY-NC), which permits unrestricted use, distribution,and reproduction in any medium, but not for commercial gain, provided the original author and source are credited.
spellingShingle Research
Li, Q. S.
Meng, F. Y.
Zhao, Y. H.
Jin, C. L.
Tian, J.
Yi, X. J.
Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells
title Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells
title_full Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells
title_fullStr Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells
title_full_unstemmed Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells
title_short Inhibition of microRNA-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type IV alpha 1 in osteoblastic MC3T3-E1 cells
title_sort inhibition of microrna-214-5p promotes cell survival and extracellular matrix formation by targeting collagen type iv alpha 1 in osteoblastic mc3t3-e1 cells
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5579316/
https://www.ncbi.nlm.nih.gov/pubmed/28784704
http://dx.doi.org/10.1302/2046-3758.68.BJR-2016-0208.R2
work_keys_str_mv AT liqs inhibitionofmicrorna2145ppromotescellsurvivalandextracellularmatrixformationbytargetingcollagentypeivalpha1inosteoblasticmc3t3e1cells
AT mengfy inhibitionofmicrorna2145ppromotescellsurvivalandextracellularmatrixformationbytargetingcollagentypeivalpha1inosteoblasticmc3t3e1cells
AT zhaoyh inhibitionofmicrorna2145ppromotescellsurvivalandextracellularmatrixformationbytargetingcollagentypeivalpha1inosteoblasticmc3t3e1cells
AT jincl inhibitionofmicrorna2145ppromotescellsurvivalandextracellularmatrixformationbytargetingcollagentypeivalpha1inosteoblasticmc3t3e1cells
AT tianj inhibitionofmicrorna2145ppromotescellsurvivalandextracellularmatrixformationbytargetingcollagentypeivalpha1inosteoblasticmc3t3e1cells
AT yixj inhibitionofmicrorna2145ppromotescellsurvivalandextracellularmatrixformationbytargetingcollagentypeivalpha1inosteoblasticmc3t3e1cells

Ejemplares similares