Cargando…

MicroRNA-455-3p regulates proliferation and osteoclast differentiation of RAW264.7 cells by targeting PTEN

BACKGROUND: Macrophages are one of the important cells in immune system. In this article, we aim to explore the regulatory role of miR-455-3p on proliferation and osteoblast differentiation of RAW264.7 cells. METHODS: Expression levels of genes and proteins in cells were tested via qRT-PCR and weste...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhang, Xiaolu, Wang, Liangming, Huang, Nianlai, Zheng, Yiqiang, Cai, Liquan, Ke, Qingfeng, Wu, Shiqiang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8994399/
https://www.ncbi.nlm.nih.gov/pubmed/35397519
http://dx.doi.org/10.1186/s12891-022-05266-0
_version_ 1784684100519460864
author Zhang, Xiaolu
Wang, Liangming
Huang, Nianlai
Zheng, Yiqiang
Cai, Liquan
Ke, Qingfeng
Wu, Shiqiang
author_facet Zhang, Xiaolu
Wang, Liangming
Huang, Nianlai
Zheng, Yiqiang
Cai, Liquan
Ke, Qingfeng
Wu, Shiqiang
author_sort Zhang, Xiaolu
collection PubMed
description BACKGROUND: Macrophages are one of the important cells in immune system. In this article, we aim to explore the regulatory role of miR-455-3p on proliferation and osteoblast differentiation of RAW264.7 cells. METHODS: Expression levels of genes and proteins in cells were tested via qRT-PCR and western blot. The targeted correlation between miR-455-3p and PTEN was identified by luciferase analysis. MTT assay and flow cytometry were applied to detect the proliferation and apoptosis of cells. Osteoclastogenesis was completed by stimulating RAW 264.7 cells with RANKL. Tartrate-resistant acid phosphatase (TRAP) activity in different groups of cells were assessed. RESULTS: Firstly, we determined that up-regulation of miR-455-3p promoted the proliferation and inhibited apoptosis of RAW 264.7 cells. MiR-455-3p deficiency played opposite effect in RAW 264.7 cells. Additionally, osteoclastogenesis-related factors (TRAP, CTSK and NFATc1) expression levels were remarkably up-regulated in miR-455-3p-mimic group of RAW264.7 cells treated with RANKL, but decreased in inhibitor group. Luciferase assay proved that miR-455-3p targeted PTEN. We took a further step and found overexpression of PTEN significantly inhibited the increased proliferation and osteoblast differentiation of RAW264.7 cells induced by miR-455-3p. CONCLUSIONS: Our findings supported basic to explore the molecular mechanism of proliferation and osteoblast differentiation of RAW264.7 cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12891-022-05266-0.
format Online
Article
Text
id pubmed-8994399
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-89943992022-04-10 MicroRNA-455-3p regulates proliferation and osteoclast differentiation of RAW264.7 cells by targeting PTEN Zhang, Xiaolu Wang, Liangming Huang, Nianlai Zheng, Yiqiang Cai, Liquan Ke, Qingfeng Wu, Shiqiang BMC Musculoskelet Disord Research BACKGROUND: Macrophages are one of the important cells in immune system. In this article, we aim to explore the regulatory role of miR-455-3p on proliferation and osteoblast differentiation of RAW264.7 cells. METHODS: Expression levels of genes and proteins in cells were tested via qRT-PCR and western blot. The targeted correlation between miR-455-3p and PTEN was identified by luciferase analysis. MTT assay and flow cytometry were applied to detect the proliferation and apoptosis of cells. Osteoclastogenesis was completed by stimulating RAW 264.7 cells with RANKL. Tartrate-resistant acid phosphatase (TRAP) activity in different groups of cells were assessed. RESULTS: Firstly, we determined that up-regulation of miR-455-3p promoted the proliferation and inhibited apoptosis of RAW 264.7 cells. MiR-455-3p deficiency played opposite effect in RAW 264.7 cells. Additionally, osteoclastogenesis-related factors (TRAP, CTSK and NFATc1) expression levels were remarkably up-regulated in miR-455-3p-mimic group of RAW264.7 cells treated with RANKL, but decreased in inhibitor group. Luciferase assay proved that miR-455-3p targeted PTEN. We took a further step and found overexpression of PTEN significantly inhibited the increased proliferation and osteoblast differentiation of RAW264.7 cells induced by miR-455-3p. CONCLUSIONS: Our findings supported basic to explore the molecular mechanism of proliferation and osteoblast differentiation of RAW264.7 cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12891-022-05266-0. BioMed Central 2022-04-09 /pmc/articles/PMC8994399/ /pubmed/35397519 http://dx.doi.org/10.1186/s12891-022-05266-0 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Zhang, Xiaolu
Wang, Liangming
Huang, Nianlai
Zheng, Yiqiang
Cai, Liquan
Ke, Qingfeng
Wu, Shiqiang
MicroRNA-455-3p regulates proliferation and osteoclast differentiation of RAW264.7 cells by targeting PTEN
title MicroRNA-455-3p regulates proliferation and osteoclast differentiation of RAW264.7 cells by targeting PTEN
title_full MicroRNA-455-3p regulates proliferation and osteoclast differentiation of RAW264.7 cells by targeting PTEN
title_fullStr MicroRNA-455-3p regulates proliferation and osteoclast differentiation of RAW264.7 cells by targeting PTEN
title_full_unstemmed MicroRNA-455-3p regulates proliferation and osteoclast differentiation of RAW264.7 cells by targeting PTEN
title_short MicroRNA-455-3p regulates proliferation and osteoclast differentiation of RAW264.7 cells by targeting PTEN
title_sort microrna-455-3p regulates proliferation and osteoclast differentiation of raw264.7 cells by targeting pten
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8994399/
https://www.ncbi.nlm.nih.gov/pubmed/35397519
http://dx.doi.org/10.1186/s12891-022-05266-0
work_keys_str_mv AT zhangxiaolu microrna4553pregulatesproliferationandosteoclastdifferentiationofraw2647cellsbytargetingpten
AT wangliangming microrna4553pregulatesproliferationandosteoclastdifferentiationofraw2647cellsbytargetingpten
AT huangnianlai microrna4553pregulatesproliferationandosteoclastdifferentiationofraw2647cellsbytargetingpten
AT zhengyiqiang microrna4553pregulatesproliferationandosteoclastdifferentiationofraw2647cellsbytargetingpten
AT cailiquan microrna4553pregulatesproliferationandosteoclastdifferentiationofraw2647cellsbytargetingpten
AT keqingfeng microrna4553pregulatesproliferationandosteoclastdifferentiationofraw2647cellsbytargetingpten
AT wushiqiang microrna4553pregulatesproliferationandosteoclastdifferentiationofraw2647cellsbytargetingpten