MiR-203 and MiR-320 Regulate Bone Morphogenetic Protein-2-Induced Osteoblast Differentiation by Targeting Distal-Less Homeobox 5 (Dlx5)

Overview

Journal Genes (Basel)

Publisher MDPI

Date 2016 Dec 28

PMID 28025541

Citations 16

Authors

Navya Laxman

Hans Mallmin

Olle Nilsson

Andreas Kindmark

Affiliations

Soon will be listed here.

Abstract

MicroRNAs (miRNAs) are a family of small, non-coding RNAs (17-24 nucleotides), which regulate gene expression either by the degradation of the target mRNAs or inhibiting the translation of genes. Recent studies have indicated that miRNA plays an important role in regulating osteoblast differentiation. In this study, we identified miR-203 and miR-320b as important miRNAs modulating osteoblast differentiation. We identified as potential common target by prediction algorithms and confirmed this by knock-down and over expression of the miRNAs and assessing at mRNA and protein levels and specificity was verified by luciferase reporter assays. We examined the effect of miR-203 and miR-320b on osteoblast differentiation by transfecting with pre- and anti-miRs. Over-expression of miR-203 and miR-320b inhibited osteoblast differentiation, whereas inhibition of miR-203 and miR-320b stimulated alkaline phosphatase activity and matrix mineralization. We show that miR-203 and miR-320b negatively regulate BMP-2-induced osteoblast differentiation by suppressing , which in turn suppresses the downstream osteogenic master transcription factor and and together they suppress osteoblast differentiation. Taken together, we propose a role for miR-203 and miR-320b in modulating bone metabolism.

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References

Chen G, Deng C, Li Y . TGF-β and BMP signaling in osteoblast differentiation and bone formation. Int J Biol Sci. 2012; 8(2):272-88. PMC: 3269610. DOI: 10.7150/ijbs.2929. View

Lee M, Kim Y, Park H, Kang A, Kyung H, Sung J . BMP-2-induced Runx2 expression is mediated by Dlx5, and TGF-beta 1 opposes the BMP-2-induced osteoblast differentiation by suppression of Dlx5 expression. J Biol Chem. 2003; 278(36):34387-94. DOI: 10.1074/jbc.M211386200. View

Groeneveld E, Burger E . Bone morphogenetic proteins in human bone regeneration. Eur J Endocrinol. 2000; 142(1):9-21. DOI: 10.1530/eje.0.1420009. View

Davis B, Hilyard A, Nguyen P, Lagna G, Hata A . Smad proteins bind a conserved RNA sequence to promote microRNA maturation by Drosha. Mol Cell. 2010; 39(3):373-84. PMC: 2921543. DOI: 10.1016/j.molcel.2010.07.011. View

Davis B, Hilyard A, Lagna G, Hata A . SMAD proteins control DROSHA-mediated microRNA maturation. Nature. 2008; 454(7200):56-61. PMC: 2653422. DOI: 10.1038/nature07086. View

Heldin C, Miyazono K, Ten Dijke P . TGF-beta signalling from cell membrane to nucleus through SMAD proteins. Nature. 1997; 390(6659):465-71. DOI: 10.1038/37284. View

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

Ulsamer A, Ortuno M, Ruiz S, Susperregui A, Osses N, Rosa J . BMP-2 induces Osterix expression through up-regulation of Dlx5 and its phosphorylation by p38. J Biol Chem. 2007; 283(7):3816-26. DOI: 10.1074/jbc.M704724200. View

Itoh T, Nozawa Y, Akao Y . MicroRNA-141 and -200a are involved in bone morphogenetic protein-2-induced mouse pre-osteoblast differentiation by targeting distal-less homeobox 5. J Biol Chem. 2009; 284(29):19272-9. PMC: 2740552. DOI: 10.1074/jbc.M109.014001. View

10.

Akiyama S, Katagiri T, Namiki M, Yamaji N, Yamamoto N, Miyama K . Constitutively active BMP type I receptors transduce BMP-2 signals without the ligand in C2C12 myoblasts. Exp Cell Res. 1997; 235(2):362-9. DOI: 10.1006/excr.1997.3680. View

11.

Stefani G, Slack F . Small non-coding RNAs in animal development. Nat Rev Mol Cell Biol. 2008; 9(3):219-30. DOI: 10.1038/nrm2347. View

12.

Hu R, Li H, Liu W, Yang L, Tan Y, Luo X . Targeting miRNAs in osteoblast differentiation and bone formation. Expert Opin Ther Targets. 2010; 14(10):1109-20. DOI: 10.1517/14728222.2010.512916. View

13.

Luo T, Lim J, Sargent T . Differential regulation of Dlx gene expression by a BMP morphogenetic gradient. Int J Dev Biol. 2001; 45(4):681-4. View

14.

Laxman N, Rubin C, Mallmin H, Nilsson O, Pastinen T, Grundberg E . Global miRNA expression and correlation with mRNA levels in primary human bone cells. RNA. 2015; 21(8):1433-43. PMC: 4509933. DOI: 10.1261/rna.049148.114. View

15.

Itoh T, Takeda S, Akao Y . MicroRNA-208 modulates BMP-2-stimulated mouse preosteoblast differentiation by directly targeting V-ets erythroblastosis virus E26 oncogene homolog 1. J Biol Chem. 2010; 285(36):27745-52. PMC: 2934642. DOI: 10.1074/jbc.M110.105080. View

16.

Grundberg E, Kwan T, Ge B, C L Lam K, Koka V, Kindmark A . Population genomics in a disease targeted primary cell model. Genome Res. 2009; 19(11):1942-52. PMC: 2775606. DOI: 10.1101/gr.095224.109. View

17.

Doench J, Sharp P . Specificity of microRNA target selection in translational repression. Genes Dev. 2004; 18(5):504-11. PMC: 374233. DOI: 10.1101/gad.1184404. View

18.

Taipaleenmaki H, Browne G, Akech J, Zustin J, van Wijnen A, Stein J . Targeting of Runx2 by miR-135 and miR-203 Impairs Progression of Breast Cancer and Metastatic Bone Disease. Cancer Res. 2015; 75(7):1433-44. PMC: 4383679. DOI: 10.1158/0008-5472.CAN-14-1026. View

19.

Krek A, Grun D, Poy M, Wolf R, Rosenberg L, Epstein E . Combinatorial microRNA target predictions. Nat Genet. 2005; 37(5):495-500. DOI: 10.1038/ng1536. View

20.

Miyama K, Yamada G, Yamamoto T, Takagi C, Miyado K, Sakai M . A BMP-inducible gene, dlx5, regulates osteoblast differentiation and mesoderm induction. Dev Biol. 1999; 208(1):123-33. DOI: 10.1006/dbio.1998.9197. View