Unveiling Novel Genes Upregulated by Both RhBMP2 and RhBMP7 During Early Osteoblastic Transdifferentiation of C2C12 Cells

Overview

Journal BMC Res Notes

Publisher Biomed Central

Specialties Biology
General Medicine

Date 2011 Sep 28

PMID 21943021

Citations 10

Authors

Juan C Bustos-Valenzuela

Andre Fujita

Erik Halcsik

Jose M Granjeiro

Mari C Sogayar

Affiliations

Soon will be listed here.

Abstract

Findings: We set out to analyse the gene expression profile of pre-osteoblastic C2C12 cells during osteodifferentiation induced by both rhBMP2 and rhBMP7 using DNA microarrays. Induced and repressed genes were intercepted, resulting in 1,318 induced genes and 704 repressed genes by both rhBMP2 and rhBMP7. We selected and validated, by RT-qPCR, 24 genes which were upregulated by rhBMP2 and rhBMP7; of these, 13 are related to transcription (Runx2, Dlx1, Dlx2, Dlx5, Id1, Id2, Id3, Fkhr1, Osx, Hoxc8, Glis1, Glis3 and Cfdp1), four are associated with cell signalling pathways (Lrp6, Dvl1, Ecsit and PKCδ) and seven are associated with the extracellular matrix (Ltbp2, Grn, Postn, Plod1, BMP1, Htra1 and IGFBP-rP10). The novel identified genes include: Hoxc8, Glis1, Glis3, Ecsit, PKCδ, LrP6, Dvl1, Grn, BMP1, Ltbp2, Plod1, Htra1 and IGFBP-rP10.

Background: BMPs (bone morphogenetic proteins) are members of the TGFβ (transforming growth factor-β) super-family of proteins, which regulate growth and differentiation of different cell types in various tissues, and play a critical role in the differentiation of mesenchymal cells into osteoblasts. In particular, rhBMP2 and rhBMP7 promote osteoinduction in vitro and in vivo, and both proteins are therapeutically applied in orthopaedics and dentistry.

Conclusion: Using DNA microarrays and RT-qPCR, we identified both previously known and novel genes which are upregulated by rhBMP2 and rhBMP7 during the onset of osteoblastic transdifferentiation of pre-myoblastic C2C12 cells. Subsequent studies of these genes in C2C12 and mesenchymal or pre-osteoblastic cells should reveal more details about their role during this type of cellular differentiation induced by BMP2 or BMP7. These studies are relevant to better understanding the molecular mechanisms underlying osteoblastic differentiation and bone repair.

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References

Moustakas A, Heldin C . Ecsit-ement on the crossroads of Toll and BMP signal transduction. Genes Dev. 2003; 17(23):2855-9. DOI: 10.1101/gad.1161403. View

Diekwisch T, Marches F, Williams A, Luan X . Cloning, gene expression, and characterization of CP27, a novel gene in mouse embryogenesis. Gene. 1999; 235(1-2):19-30. DOI: 10.1016/s0378-1119(99)00220-6. View

Tu X, Joeng K, Nakayama K, Nakayama K, Rajagopal J, Carroll T . Noncanonical Wnt signaling through G protein-linked PKCdelta activation promotes bone formation. Dev Cell. 2007; 12(1):113-27. PMC: 1861818. DOI: 10.1016/j.devcel.2006.11.003. View

Osyczka A, Diefenderfer D, Bhargave G, Leboy P . Different effects of BMP-2 on marrow stromal cells from human and rat bone. Cells Tissues Organs. 2004; 176(1-3):109-19. PMC: 1463182. DOI: 10.1159/000075032. View

Hollnagel A, Oehlmann V, Heymer J, Ruther U, Nordheim A . Id genes are direct targets of bone morphogenetic protein induction in embryonic stem cells. J Biol Chem. 1999; 274(28):19838-45. DOI: 10.1074/jbc.274.28.19838. View

Bois P, Grosveld G . FKHR (FOXO1a) is required for myotube fusion of primary mouse myoblasts. EMBO J. 2003; 22(5):1147-57. PMC: 150349. DOI: 10.1093/emboj/cdg116. View

Deschaseaux F, Sensebe L, Heymann D . Mechanisms of bone repair and regeneration. Trends Mol Med. 2009; 15(9):417-29. DOI: 10.1016/j.molmed.2009.07.002. View

Wharton Jr K . Runnin' with the Dvl: proteins that associate with Dsh/Dvl and their significance to Wnt signal transduction. Dev Biol. 2002; 253(1):1-17. DOI: 10.1006/dbio.2002.0869. View

Gimble J, Morgan C, Kelly K, Wu X, Dandapani V, Wang C . Bone morphogenetic proteins inhibit adipocyte differentiation by bone marrow stromal cells. J Cell Biochem. 1995; 58(3):393-402. DOI: 10.1002/jcb.240580312. View

10.

Uzawa K, Grzesik W, Nishiura T, Kuznetsov S, Robey P, Brenner D . Differential expression of human lysyl hydroxylase genes, lysine hydroxylation, and cross-linking of type I collagen during osteoblastic differentiation in vitro. J Bone Miner Res. 1999; 14(8):1272-80. DOI: 10.1359/jbmr.1999.14.8.1272. View

11.

Siqueira M, Flowers S, Bhattacharya R, Faibish D, Behl Y, Kotton D . FOXO1 modulates osteoblast differentiation. Bone. 2011; 48(5):1043-51. PMC: 3109483. DOI: 10.1016/j.bone.2011.01.019. View

12.

Mbalaviele G, Sheikh S, Stains J, Salazar V, Cheng S, Chen D . Beta-catenin and BMP-2 synergize to promote osteoblast differentiation and new bone formation. J Cell Biochem. 2004; 94(2):403-18. PMC: 2647989. DOI: 10.1002/jcb.20253. View

13.

Katagiri T, Yamaguchi A, Komaki M, Abe E, Takahashi N, Ikeda T . Bone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage. J Cell Biol. 1994; 127(6 Pt 1):1755-66. PMC: 2120318. DOI: 10.1083/jcb.127.6.1755. View

14.

Newton A . Regulation of protein kinase C. Curr Opin Cell Biol. 1997; 9(2):161-7. DOI: 10.1016/s0955-0674(97)80058-0. View

15.

Kokubu C, Heinzmann U, Kokubu T, Sakai N, Kubota T, Kawai M . Skeletal defects in ringelschwanz mutant mice reveal that Lrp6 is required for proper somitogenesis and osteogenesis. Development. 2004; 131(21):5469-80. DOI: 10.1242/dev.01405. View

16.

Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A . Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002; 3(7):RESEARCH0034. PMC: 126239. DOI: 10.1186/gb-2002-3-7-research0034. View

17.

Hassan M, Tare R, Lee S, Mandeville M, Morasso M, Javed A . BMP2 commitment to the osteogenic lineage involves activation of Runx2 by DLX3 and a homeodomain transcriptional network. J Biol Chem. 2006; 281(52):40515-26. DOI: 10.1074/jbc.M604508200. View

18.

Miyazono K . Signal transduction by bone morphogenetic protein receptors: functional roles of Smad proteins. Bone. 1999; 25(1):91-3. DOI: 10.1016/s8756-3282(99)00113-1. View

19.

Gaur T, Lengner C, Hovhannisyan H, Bhat R, Bodine P, Komm B . Canonical WNT signaling promotes osteogenesis by directly stimulating Runx2 gene expression. J Biol Chem. 2005; 280(39):33132-40. DOI: 10.1074/jbc.M500608200. View

20.

Balint E, Lapointe D, Drissi H, van der Meijden C, Young D, van Wijnen A . Phenotype discovery by gene expression profiling: mapping of biological processes linked to BMP-2-mediated osteoblast differentiation. J Cell Biochem. 2003; 89(2):401-26. DOI: 10.1002/jcb.10515. View