miR-26b-5p/TCF-4 Controls the Adipogenic Differentiation of Human Adipose-derived Mesenchymal Stem Cells

In this study, we assessed the ability of miR-26b-5p to regulate T cell factor 4 (TCF-4) expression and thereby control human adipose-derived mesenchymal stem cell (hADMSC) adipogenic differentiation. Adipogenic medium was used to induce hADMSC differentiation over a 6-d period. The ability of miR-2...

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Main Authors: Yadong Luo, Huan Ji, Yan Cao, Xu Ding, Meng Li, Haiyang Song, Sheng Li, Chenxing WaTableng, Heming Wu MD, Jian Meng, Hongming Du
Format: Article
Language:English
Published: SAGE Publishing 2020-06-01
Series:Cell Transplantation
Online Access:https://doi.org/10.1177/0963689720934418
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author Yadong Luo
Huan Ji
Yan Cao
Xu Ding
Meng Li
Haiyang Song
Sheng Li
Chenxing WaTableng
Heming Wu MD
Jian Meng
Hongming Du
author_facet Yadong Luo
Huan Ji
Yan Cao
Xu Ding
Meng Li
Haiyang Song
Sheng Li
Chenxing WaTableng
Heming Wu MD
Jian Meng
Hongming Du
author_sort Yadong Luo
collection DOAJ
description In this study, we assessed the ability of miR-26b-5p to regulate T cell factor 4 (TCF-4) expression and thereby control human adipose-derived mesenchymal stem cell (hADMSC) adipogenic differentiation. Adipogenic medium was used to induce hADMSC differentiation over a 6-d period. The ability of miR-26b-5p to interact with the TCF-4 mRNA was confirmed through both predictive bioinformatics analyses and luciferase reporter assays. Immunofluorescent staining was used to visualize the impact of miR-26b-5p inhibition or overexpression on TCF-4 and β-catenin levels in hADMSCs. Further functional analyses were conducted by transfecting these cells with siRNAs specific for TCF-4 and β-catenin. Adipogenic marker and Wnt/β-catenin pathway gene expression levels were assessed via real-time polymerase chain reaction and western blotting. β-catenin localization was assessed via immunofluorescent staining. As expected, our adipogenic media induced the adipocytic differentiation of hADMSCs. In addition, we confirmed that TCF-4 is an miR-26b-5p target gene in these cells, and that protein levels of both TCF-4 and β-catenin were reduced when these cells were transfected with miR-26b-5p mimics. Overexpression of this microRNA also enhanced hADMSC adipogenesis, whereas TCF-4 and β-catenin overexpression inhibited this process. The enhanced hADMSC adipogenic differentiation that was observed following TCF-4 or β-catenin knockdown was partially reversed when miR-26b-5p expression was inhibited. We found that miR-26b-5p serves as a direct negative regulator of TCF-4 expression within hADMSCs, leading to inactivation of the Wnt/β-catenin pathway and thereby promoting the adipogenic differentiation of these cells in vitro .
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spelling doaj.art-dc50bca992ee4447abe0973cc81dad302022-12-22T01:18:20ZengSAGE PublishingCell Transplantation1555-38922020-06-012910.1177/0963689720934418miR-26b-5p/TCF-4 Controls the Adipogenic Differentiation of Human Adipose-derived Mesenchymal Stem CellsYadong Luo0Huan Ji1Yan Cao2Xu Ding3Meng Li4Haiyang Song5Sheng Li6Chenxing WaTableng7Heming Wu MD8Jian Meng9Hongming Du10 These authors contributed equally to this article These authors contributed equally to this article These authors contributed equally to this article Jiangsu Key Laboratory of Oral Disease, Nanjing Medical University, Nanjing, Jiangsu Province, PR China Jiangsu Key Laboratory of Oral Disease, Nanjing Medical University, Nanjing, Jiangsu Province, PR China Jiangsu Key Laboratory of Oral Disease, Nanjing Medical University, Nanjing, Jiangsu Province, PR China Jiangsu Key Laboratory of Oral Disease, Nanjing Medical University, Nanjing, Jiangsu Province, PR China Jiangsu Key Laboratory of Oral Disease, Nanjing Medical University, Nanjing, Jiangsu Province, PR China Jiangsu Key Laboratory of Oral Disease, Nanjing Medical University, Nanjing, Jiangsu Province, PR China Both authors are co-corresponding authors Both authors are co-corresponding authorsIn this study, we assessed the ability of miR-26b-5p to regulate T cell factor 4 (TCF-4) expression and thereby control human adipose-derived mesenchymal stem cell (hADMSC) adipogenic differentiation. Adipogenic medium was used to induce hADMSC differentiation over a 6-d period. The ability of miR-26b-5p to interact with the TCF-4 mRNA was confirmed through both predictive bioinformatics analyses and luciferase reporter assays. Immunofluorescent staining was used to visualize the impact of miR-26b-5p inhibition or overexpression on TCF-4 and β-catenin levels in hADMSCs. Further functional analyses were conducted by transfecting these cells with siRNAs specific for TCF-4 and β-catenin. Adipogenic marker and Wnt/β-catenin pathway gene expression levels were assessed via real-time polymerase chain reaction and western blotting. β-catenin localization was assessed via immunofluorescent staining. As expected, our adipogenic media induced the adipocytic differentiation of hADMSCs. In addition, we confirmed that TCF-4 is an miR-26b-5p target gene in these cells, and that protein levels of both TCF-4 and β-catenin were reduced when these cells were transfected with miR-26b-5p mimics. Overexpression of this microRNA also enhanced hADMSC adipogenesis, whereas TCF-4 and β-catenin overexpression inhibited this process. The enhanced hADMSC adipogenic differentiation that was observed following TCF-4 or β-catenin knockdown was partially reversed when miR-26b-5p expression was inhibited. We found that miR-26b-5p serves as a direct negative regulator of TCF-4 expression within hADMSCs, leading to inactivation of the Wnt/β-catenin pathway and thereby promoting the adipogenic differentiation of these cells in vitro .https://doi.org/10.1177/0963689720934418
spellingShingle Yadong Luo
Huan Ji
Yan Cao
Xu Ding
Meng Li
Haiyang Song
Sheng Li
Chenxing WaTableng
Heming Wu MD
Jian Meng
Hongming Du
miR-26b-5p/TCF-4 Controls the Adipogenic Differentiation of Human Adipose-derived Mesenchymal Stem Cells
Cell Transplantation
title miR-26b-5p/TCF-4 Controls the Adipogenic Differentiation of Human Adipose-derived Mesenchymal Stem Cells
title_full miR-26b-5p/TCF-4 Controls the Adipogenic Differentiation of Human Adipose-derived Mesenchymal Stem Cells
title_fullStr miR-26b-5p/TCF-4 Controls the Adipogenic Differentiation of Human Adipose-derived Mesenchymal Stem Cells
title_full_unstemmed miR-26b-5p/TCF-4 Controls the Adipogenic Differentiation of Human Adipose-derived Mesenchymal Stem Cells
title_short miR-26b-5p/TCF-4 Controls the Adipogenic Differentiation of Human Adipose-derived Mesenchymal Stem Cells
title_sort mir 26b 5p tcf 4 controls the adipogenic differentiation of human adipose derived mesenchymal stem cells
url https://doi.org/10.1177/0963689720934418
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