Sox9-Increased miR-322-5p Facilitates BMP2-Induced Chondrogenic Differentiation by Targeting Smad7 in Mesenchymal Stem Cells

Bone morphogenetic protein 2 (BMP2) induces effective chondrogenesis of mesenchymal stem cells (MSCs) by promoting Sox9 expression. However, BMP2 also induces chondrocyte hypertrophy and endochondral ossification by upregulating Smad7 expression, which leads to the disruption of chondrogenesis. In a...

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Autores principales:	Yongsheng Zeng, Chengcheng Du, Pengcheng Xiao, Yiting Lei, Piao Zhao, Zhenglin Zhu, Shengqiang Gao, Bowen Chen, Shengwen Cheng, Wei Huang, Chen Zhao
Formato:	article
Lenguaje:	EN
Publicado:	Hindawi Limited 2021
Materias:	Internal medicine RC31-1245
Acceso en línea:	https://doaj.org/article/26fd36b2fa71447b88c01eecc8731514
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spelling	oai:doaj.org-article:26fd36b2fa71447b88c01eecc87315142021-11-15T01:19:30ZSox9-Increased miR-322-5p Facilitates BMP2-Induced Chondrogenic Differentiation by Targeting Smad7 in Mesenchymal Stem Cells1687-967810.1155/2021/9778207https://doaj.org/article/26fd36b2fa71447b88c01eecc87315142021-01-01T00:00:00Zhttp://dx.doi.org/10.1155/2021/9778207https://doaj.org/toc/1687-9678Bone morphogenetic protein 2 (BMP2) induces effective chondrogenesis of mesenchymal stem cells (MSCs) by promoting Sox9 expression. However, BMP2 also induces chondrocyte hypertrophy and endochondral ossification by upregulating Smad7 expression, which leads to the disruption of chondrogenesis. In addition, Smad7 can be inhibited by Sox9. Therefore, the underlying mechanism is not clear. Currently, an increasing number of studies have shown that microRNAs play a pivotal role in chondrogenic and pathophysiological processes of cartilage. The purpose of this study was to determine which microRNA is increased by Sox9 and targets Smad7, thus assisting BMP2 in maintaining stable chondrogenesis. We found that miR-322-5p meets the requirement through next-generation sequencing (NGS) and bioinformatic analysis. The targeting relationship between miR-322-5p and Smad7 was confirmed by dual-luciferase reporter assays, qPCR, and western blotting (WB). The in vitro study indicated that overexpression of miR-322-5p significantly inhibited Smad7 expression, thus causing increased chondrogenic differentiation and decreased hypertrophic differentiation, while silencing of miR-322-5p led to the opposite results. Flow cytometry (FCM) analysis indicated that overexpression of miR-322-5p significantly decreased the rate of early apoptosis in BMP2-stimulated MSCs, while silencing of miR-322-5p increased the rate. A mouse limb explant assay revealed that the expression of miR-322-5p was negatively correlated with the length of the BMP2-stimulated hypertrophic zone of the growth plate. An in vivo study also confirmed that miR-322-5p assisted BMP2 in chondrogenic differentiation. Taken together, our results suggested that Sox9-increased miR-322-5p expression can promote BMP2-induced chondrogenesis by targeting Smad7, which can be exploited for effective tissue engineering of cartilage.Yongsheng ZengChengcheng DuPengcheng XiaoYiting LeiPiao ZhaoZhenglin ZhuShengqiang GaoBowen ChenShengwen ChengWei HuangChen ZhaoHindawi LimitedarticleInternal medicineRC31-1245ENStem Cells International, Vol 2021 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Internal medicine RC31-1245
spellingShingle	Internal medicine RC31-1245 Yongsheng Zeng Chengcheng Du Pengcheng Xiao Yiting Lei Piao Zhao Zhenglin Zhu Shengqiang Gao Bowen Chen Shengwen Cheng Wei Huang Chen Zhao Sox9-Increased miR-322-5p Facilitates BMP2-Induced Chondrogenic Differentiation by Targeting Smad7 in Mesenchymal Stem Cells
description	Bone morphogenetic protein 2 (BMP2) induces effective chondrogenesis of mesenchymal stem cells (MSCs) by promoting Sox9 expression. However, BMP2 also induces chondrocyte hypertrophy and endochondral ossification by upregulating Smad7 expression, which leads to the disruption of chondrogenesis. In addition, Smad7 can be inhibited by Sox9. Therefore, the underlying mechanism is not clear. Currently, an increasing number of studies have shown that microRNAs play a pivotal role in chondrogenic and pathophysiological processes of cartilage. The purpose of this study was to determine which microRNA is increased by Sox9 and targets Smad7, thus assisting BMP2 in maintaining stable chondrogenesis. We found that miR-322-5p meets the requirement through next-generation sequencing (NGS) and bioinformatic analysis. The targeting relationship between miR-322-5p and Smad7 was confirmed by dual-luciferase reporter assays, qPCR, and western blotting (WB). The in vitro study indicated that overexpression of miR-322-5p significantly inhibited Smad7 expression, thus causing increased chondrogenic differentiation and decreased hypertrophic differentiation, while silencing of miR-322-5p led to the opposite results. Flow cytometry (FCM) analysis indicated that overexpression of miR-322-5p significantly decreased the rate of early apoptosis in BMP2-stimulated MSCs, while silencing of miR-322-5p increased the rate. A mouse limb explant assay revealed that the expression of miR-322-5p was negatively correlated with the length of the BMP2-stimulated hypertrophic zone of the growth plate. An in vivo study also confirmed that miR-322-5p assisted BMP2 in chondrogenic differentiation. Taken together, our results suggested that Sox9-increased miR-322-5p expression can promote BMP2-induced chondrogenesis by targeting Smad7, which can be exploited for effective tissue engineering of cartilage.
format	article
author	Yongsheng Zeng Chengcheng Du Pengcheng Xiao Yiting Lei Piao Zhao Zhenglin Zhu Shengqiang Gao Bowen Chen Shengwen Cheng Wei Huang Chen Zhao
author_facet	Yongsheng Zeng Chengcheng Du Pengcheng Xiao Yiting Lei Piao Zhao Zhenglin Zhu Shengqiang Gao Bowen Chen Shengwen Cheng Wei Huang Chen Zhao
author_sort	Yongsheng Zeng
title	Sox9-Increased miR-322-5p Facilitates BMP2-Induced Chondrogenic Differentiation by Targeting Smad7 in Mesenchymal Stem Cells
title_short	Sox9-Increased miR-322-5p Facilitates BMP2-Induced Chondrogenic Differentiation by Targeting Smad7 in Mesenchymal Stem Cells
title_full	Sox9-Increased miR-322-5p Facilitates BMP2-Induced Chondrogenic Differentiation by Targeting Smad7 in Mesenchymal Stem Cells
title_fullStr	Sox9-Increased miR-322-5p Facilitates BMP2-Induced Chondrogenic Differentiation by Targeting Smad7 in Mesenchymal Stem Cells
title_full_unstemmed	Sox9-Increased miR-322-5p Facilitates BMP2-Induced Chondrogenic Differentiation by Targeting Smad7 in Mesenchymal Stem Cells
title_sort	sox9-increased mir-322-5p facilitates bmp2-induced chondrogenic differentiation by targeting smad7 in mesenchymal stem cells
publisher	Hindawi Limited
publishDate	2021
url	https://doaj.org/article/26fd36b2fa71447b88c01eecc8731514
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Sox9-Increased miR-322-5p Facilitates BMP2-Induced Chondrogenic Differentiation by Targeting Smad7 in Mesenchymal Stem Cells

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