YTHDC2 inhibits rat bone mesenchymal stem cells osteogenic differentiation by accelerating RUNX2 mRNA degradation via m6A methylation

Bo Ma; Pei Cao; Lichen Zhang; Hongyi Zhu; Xuwen Ye; Lingjun Wang; Liang Chen

doi:10.1016/j.heliyon.2023.e18876

YTHDC2 inhibits rat bone mesenchymal stem cells osteogenic differentiation by accelerating RUNX2 mRNA degradation via m6A methylation

Heliyon. 2023 Aug 4;9(8):e18876. doi: 10.1016/j.heliyon.2023.e18876. eCollection 2023 Aug.

Authors

Bo Ma^{1

2}, Pei Cao^{3

4}, Lichen Zhang², Hongyi Zhu², Xuwen Ye², Lingjun Wang², Liang Chen²

Affiliations

¹ Department of Trauma and Orthopedics, Peking University People's Hospital, Beijing, PR China.
² Department of Orthopedics, The First Affiliated Hospital of Soochow University, Orthopedic Institute, Soochow University, Suzhou, Jiangsu, 215006, PR China.
³ Nankai University School of Medicine, Tianjin, PR China.
⁴ Department of General Surgery, The First Affiliated Hospital of Soochow University, Suzhou, 215006, Jiangsu Province, PR China.

Abstract

As the most abundant internal mRNA modification, N6-methyladenosine (m6A) RNA methylation has been found to influence many biological events including bone mesenchymal stem cells (BMSCs) osteogenic differentiation. YTH N6-methyladenosine RNA binding protein C2 （YTHDC2） is an m6A reading protein with the ability to mediate the decay of combined methylated mRNA, however its role in BMSCs osteogenic differentiation remains unknown. In this study, we first found an increase of RUNX family transcription factor 2 （RUNX2） expression and a decrease of YTHDC2 expression during the process of BMSCs osteogenic differentiation. Furthermore, we transfected BMSCs with YTHDC2 interference fragment, resulting in an increased content of RUNX2 mRNA and protein inside BMSCs. Finally, through RNA Immunoprecipitation experiments, we confirmed that YTHDC2 protein can bind to RUNX2 mRNA and accelerate its decomposition. Moreover, the immunofluorescence staining also showed a negative correlation between YTHDC2 and RUNX2. In conclusion, during BMSCs osteogenic differentiation, YTHDC2 protein showed decreased expression, resulting in a higher level of RUNX2 (mRNA and protein) expression inside cells, indicating YTHDC2 as a promising molecular target for the regulation of BMSCs osteogenic differentiation.

Keywords: BMSCs; Osteogenic differentiation; RUNX2; YTHDC2; m6A.