MicroRNA-204 regulates osteogenic induction in dental follicle cells

Ko Ito; Risa Tomoki; Naomi Ogura; Kosuke Takahashi; Takashi Eda; Fumie Yamazaki; Yugo Kato; Alastair Goss; Toshirou Kondoh

doi:10.1016/j.jds.2019.11.004

MicroRNA-204 regulates osteogenic induction in dental follicle cells

J Dent Sci. 2020 Dec;15(4):457-465. doi: 10.1016/j.jds.2019.11.004. Epub 2020 Jan 2.

Authors

Ko Ito^{1

2}, Risa Tomoki¹, Naomi Ogura^{1

2}, Kosuke Takahashi^{1

2}, Takashi Eda¹, Fumie Yamazaki¹, Yugo Kato¹, Alastair Goss³, Toshirou Kondoh^{1

2}

Affiliations

¹ Department of Maxillofacial Surgery, Nihon University School of Dentistry at Matsudo, Chiba, Japan.
² Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, Chiba, Japan.
³ Oral and Maxillofacial Surgery Unit, Faculty of Health Science, University of Adelaide, South Australia, Australia.

Abstract

The dental follicle is an ectomesenchymal tissue surrounding developing tooth germ that contains osteoblastic-lineage-committed stem/progenitor cells. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression during stem cell growth, proliferation, and differentiation. The aim of this study was to investigate the key regulators of miRNA during osteogenic differentiation in human dental follicle cells (hDFC). We analyzed miRNA expression profiles in hDFC during osteoblastic differentiation. Expression of miR-204 was decreased in hDFC during osteogenic induction on microarray analysis. Real-time and RT-PCR analysis also showed that the expression of miR-204 was decreased in all three hDFC during osteogenic differentiation. To investigate whether miR-204 has an effect on osteogenic differentiation, miR-204 was predicted to target alkaline phosphatase (ALP), secreted protein acidic and rich in cysteine (SPARC), and Runx2 in the in the 3'-UTRs by in silico analysis. When miR-204 was transfected into hDFC, the activity of ALP and protein levels of SPARC and Runx2 were decreased. mRNA levels of ALP, SPARC and Runx2 were also decreased by miR-204 transfection. Our data suggest that miR-204 negatively regulates the osteogenic differentiation of hDFC by targeting the bone-specific transcription factor Runx2, the mineralization maker ALP and the bone extracellular matrix protein SPARC.

Keywords: Human dental follicle; Micro RNA; MicroRNA 204; Osteogenic differentiation.