- 作者：Fan Yang ; DDS ; PhD&lowast ; ; yangfan@fmmu.edu.cn" class="auth_mail" title="E-mail the corresponding author ; Na Xu ; DDS ; MS&dagger ; ; Dongmei Li ; DDS ; PhD&Dagger ; ; Lina Guan ; DDS ; MS&lowast ; ; Ying He ; DDS ; MS&lowast ; ; Yaqing Zhang ; DDS ; PhD&lowast ; ; Qun Lu ; DDS ; PhD&lowast ; ; Xudong Zhang ; DDS ; PhD§ ;
- 关键词：Adult stem cell research ; cell differentiation ; dental pulps ; feedback ; ligases ; mesenchymal stem cells ; runt-related transcription factor 2 ; smad ubiquitylation regulatory factor-1 ; ubiquitination
- 刊名：Journal of Endodontics
- 出版年：2014
- 出版时间：October 2014
- 年：2014
- 卷：40
- 期：10
- 页码：1579-1586
- 全文大小：1173 K
Methods
RUNX2 or SMURF1 expression was silenced in hDPSCs by lentiviral transduction of short hairpin RNA . The relationship between RUNX2 and SMURF1 expression was analyzed using quantitative polymerase chain reaction, Western blotting, dual luciferase reporter assays, and chromatin immunoprecipitation. The effect of the interplay between RUNX2 and SMURF1 on the odontoblastic differentiation of hDPSCs was examined in SMURF1-deficient hDPSCs.
Results
The inhibition of SMURF1 in hDPSCs significantly increased RUNX2 at the protein level that was associated with decreased RUNX2 ubiquitination but did not affect RUNX2 messenger RNA expression. On the other hand, depletion of RUNX2 in hDPSCs decreased SMURF1 at both the protein and messenger RNA levels. A RUNX2-binding motif at −308 bp of the SMURF1 promoter functioned in RUNX2-mediated SMURF1 expression. Moreover, the expression levels of RUNX2 were associated with SMURF1 levels during odontoblastic differentiation. Significantly, the knockdown of SMURF1 up-regulated RUNX2 expression and down-regulated dentin sialophosphoprotein and dental matrix protein-1 expression in odontoblastic differentiation.
Conclusions
These results reveal the regulatory circuit between RUNX2 and SMURF1 controls RUNX2 expression and regulates odntoblastic differentiation in hDPSCs.