MicroRNA Hsa-Let-7b Regulates the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Targeting CTHRC1

Let-7 miRNA family has been proved as a key regulator of mesenchymal stem cells’ (MSCs’) biological features. However, whether let-7b could affect the differentiation or proliferation of periodontal ligament stem cells (PDLSCs) is still unknown. Here, we found that the expression of hsa-let-7b was v...

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Main Authors: Lin Fu, Na Li, Yu Ye, Xiaying Ye, Tong Xiao, Xiao Wu, Yanxia Ma, Jinhua Yu
Format: Article
Language:English
Published: Wiley 2021-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2021/5791181
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author Lin Fu
Na Li
Yu Ye
Xiaying Ye
Tong Xiao
Xiao Wu
Yanxia Ma
Jinhua Yu
author_facet Lin Fu
Na Li
Yu Ye
Xiaying Ye
Tong Xiao
Xiao Wu
Yanxia Ma
Jinhua Yu
author_sort Lin Fu
collection DOAJ
description Let-7 miRNA family has been proved as a key regulator of mesenchymal stem cells’ (MSCs’) biological features. However, whether let-7b could affect the differentiation or proliferation of periodontal ligament stem cells (PDLSCs) is still unknown. Here, we found that the expression of hsa-let-7b was visibly downregulated after mineralization induction of PDLSCs. After transfected with hsa-let-7b mimics or inhibitor reagent, the proliferation ability of PDLSCs was detected by cell counting kit-8 (CCK-8), flow cytometry, and 5-ethynyl-2-deoxyuridine (EdU) assay. On the other hand, the osteogenic differentiation capacity was detected by alkaline phosphatase (ALP) staining and activity, alizarin red staining, Western blot, and quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR). We verified that hsa-let-7b did not significantly impact the proliferation ability of PDLSCs, but it could curb the osteogenic differentiation of PDLSCs. Besides, we predicted CTHRC1 acts as the downstream gene of hsa-let-7b to affect this process. Moreover, the combination of CTHRC1 and hsa-let-7b was verified by dual luciferase reporter assay. Our results demonstrated that the osteogenic differentiation of PDLSCs was enhanced after inhibiting hsa-let-7b, while was weakened after cotransfection with Si-CTHRC1. Collectively, hsa-let-7b can repress the osteogenic differentiation of PDLSCs by regulating CTHRC1.
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spelling doaj-art-12b4ea64bdb24242a3bef7e4527391e52025-08-20T02:02:12ZengWileyStem Cells International1687-96782021-01-01202110.1155/2021/5791181MicroRNA Hsa-Let-7b Regulates the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Targeting CTHRC1Lin Fu0Na Li1Yu Ye2Xiaying Ye3Tong Xiao4Xiao Wu5Yanxia Ma6Jinhua Yu7Key Laboratory of Oral Diseases of Jiangsu Province and Stomatological Institute of Nanjing Medical UniversityKey Laboratory of Oral Diseases of Jiangsu Province and Stomatological Institute of Nanjing Medical UniversityKey Laboratory of Oral Diseases of Jiangsu Province and Stomatological Institute of Nanjing Medical UniversityKey Laboratory of Oral Diseases of Jiangsu Province and Stomatological Institute of Nanjing Medical UniversityKey Laboratory of Oral Diseases of Jiangsu Province and Stomatological Institute of Nanjing Medical UniversityKey Laboratory of Oral Diseases of Jiangsu Province and Stomatological Institute of Nanjing Medical UniversityKey Laboratory of Oral Diseases of Jiangsu Province and Stomatological Institute of Nanjing Medical UniversityKey Laboratory of Oral Diseases of Jiangsu Province and Stomatological Institute of Nanjing Medical UniversityLet-7 miRNA family has been proved as a key regulator of mesenchymal stem cells’ (MSCs’) biological features. However, whether let-7b could affect the differentiation or proliferation of periodontal ligament stem cells (PDLSCs) is still unknown. Here, we found that the expression of hsa-let-7b was visibly downregulated after mineralization induction of PDLSCs. After transfected with hsa-let-7b mimics or inhibitor reagent, the proliferation ability of PDLSCs was detected by cell counting kit-8 (CCK-8), flow cytometry, and 5-ethynyl-2-deoxyuridine (EdU) assay. On the other hand, the osteogenic differentiation capacity was detected by alkaline phosphatase (ALP) staining and activity, alizarin red staining, Western blot, and quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR). We verified that hsa-let-7b did not significantly impact the proliferation ability of PDLSCs, but it could curb the osteogenic differentiation of PDLSCs. Besides, we predicted CTHRC1 acts as the downstream gene of hsa-let-7b to affect this process. Moreover, the combination of CTHRC1 and hsa-let-7b was verified by dual luciferase reporter assay. Our results demonstrated that the osteogenic differentiation of PDLSCs was enhanced after inhibiting hsa-let-7b, while was weakened after cotransfection with Si-CTHRC1. Collectively, hsa-let-7b can repress the osteogenic differentiation of PDLSCs by regulating CTHRC1.http://dx.doi.org/10.1155/2021/5791181
spellingShingle Lin Fu
Na Li
Yu Ye
Xiaying Ye
Tong Xiao
Xiao Wu
Yanxia Ma
Jinhua Yu
MicroRNA Hsa-Let-7b Regulates the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Targeting CTHRC1
Stem Cells International
title MicroRNA Hsa-Let-7b Regulates the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Targeting CTHRC1
title_full MicroRNA Hsa-Let-7b Regulates the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Targeting CTHRC1
title_fullStr MicroRNA Hsa-Let-7b Regulates the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Targeting CTHRC1
title_full_unstemmed MicroRNA Hsa-Let-7b Regulates the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Targeting CTHRC1
title_short MicroRNA Hsa-Let-7b Regulates the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells by Targeting CTHRC1
title_sort microrna hsa let 7b regulates the osteogenic differentiation of human periodontal ligament stem cells by targeting cthrc1
url http://dx.doi.org/10.1155/2021/5791181
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