DOI: 10.3724/SP.J.1206.2012.00620

Progress in Biochemistry and Biophysics (生物化学与生物物理进展) 2013/40:12 PP.1220-1229

BMP9 Regulates Osteogenic Differentiation of Mesenchymal Stem Cells Through JNKs Kinase Pathway

In addition to Smad pathway, our previous study has shown that BMP9 can induce osteogenicdifferentiation of mesenchymal stem cells (MSCs) through p38 MAPKs pathway. In this study, we explore thepossible involvement and detail role of JNKs (c-Jun N-terminal kinases) in BMP9-indcued osteogenicdifferentiation of MSCs. BMP9 was introduced into MSCs by recombinant adenoviruses protocol, then, in vivoand in vivo assays were conducted to detect whether BMP9 can induce osteogenic differentiation of MSCs throughJNKs kinase pathway. The results showed that BMP9 can activate JNKs kinase through increase thephosphorylated form of JNKs kinase. JNKs kinase inhibitor SP600125 can inhibit ALP activity, OPN and OCNexpression, as well as calcium deposition induced by BMP9 in MSCs. Furthermore, SP600125 also led to adecrease in BMP9-induced Runx2 activity and canonical Smad signaling. Moreover, when JNKs kinase wassilenced by RNA interference in MSCs, BMP9-induced osteogenic differentiation in vivo and ectopic boneformation in vivo were accordingly inhibited along with knockdown of JNKs. Taken together, those resultsintensively suggested that BMP9 can induce and regulate osteogenic differentiation of MSCs through activatingJNKs kinase pathway.

Key words:bone morphogenetic proteins 9,JNKs,mesenchymal stem cells,osteogenic differentiation,mitogen activated protein kinase

ReleaseDate:2015-04-18 09:14:50

[1] Stappenbeck T S, Miyoshi H. The role of stromal stem cells in tissue regeneration and wound repair. Science, 2009, 324 (5935):1666-1669

[2] Myers T J, Granero-Molto F, Longobardi L, et al. Mesenchymal stem cells at the intersection of cell and gene therapy. Expert Opin Biol Ther, 2010, 10(12): 1663-1679

[3] Wu Y, Zhao R C, Tredget E E. Concise review: bone marrowderived stem/progenitor cells in cutaneous repair and regeneration. Stem Cells, 2010, 28(5): 905-915

[4] Arthur A, Zannettino A, Gronthos S. The therapeutic applications of multipotential mesenchymal/stromal stem cells in skeletal tissue repair. J Cell Physiol, 2009, 218(2): 237-245

[5] Deschaseaux F, Pontikoglou C, Sens éb é L. Bone regeneration: the stem/progenitor cells point of view. J Cell Mol Med, 2010,14(1-2): 103-115

[6] McKay W F, Peckham S M, Badura J M. A comprehensive clinical review of recombinant human bone morphogenetic protein-2 (INFUSE Bone Graft). Int Orthop, 2007, 31(6): 729-734

[7] Acil Y, Springer I N, Broek V, et al Effects of bone morphogenetic protein-7 stimulation on osteoblasts cultured on different biomaterials. J Cell Biochem, 2002, 86(1): 90-98

[8] Luu H H, Song W X, Luo X, et al. Distinct roles of bone morphogenetic proteins in osteogenic differentiation of mesenchymal stem cells. J Orthop Res, 2007, 25(5): 665-677

[9] Miller A F, Harvey S A, Thies R S, et al Bone morphogeneticprotein-9, an autocrine/paracrine cytokine in the liver. J Biol Chem, 2000, 275(24): 17937-17945

[10] Chen C, Grzegorzewski K J, Barash S, et al. An integrated functional genomics screening program reveals a role for BMP-9 in glucose homeostasis. Nat Biotechnol, 2003, 21(3): 294-301

[11] Truksa J, Peng H, Lee P, et al. Bone morphogenetic proteins 2, 4, and 9 stimulate murine hepcidin 1 expression independently of Hfe, transferring receptor 2 (Tfr2), and IL-6. Proc Natl Acad Sci USA, 2006, 103(27): 10289-10291

[12] López-Coviella I, Berse B, Krauss R, et al. Induction and maintenance of the neuronal cholinergic phenotype in the central nervous system by BMP9. Science, 2000, 289(5477): 313-316

[13] Kang Q, Sun M H, Cheng H, et al. Characterization of the distinct orthotopic bone forming activity of 14 BMPs using recombinant adenovirus-mediated gene delivery. Gene Ther, 2004, 11 (17):1312-1320

[14] 张 燕, 文 巍, 罗进勇. 骨形态发生蛋白 9 定向诱导多潜能干细 胞 成 骨 分 化 . 生 物 化 学 与 生 物 物 理 进 展 , 2009, 36 (10):1291-1298 Zhang Y, Wen W, Luo J Y. Prog Biochem Biophys, 2009, 36(10):1291-1298

[15] Cargnello M, Roux P P. Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases. MicrobiolMol Biol Rev, 2011, 75(1): 50-83

[16] N觟th U, Tuli R, Seghatoleslami R, et al. Activation of p38 and Smads mediates BMP-2 effects on human trabecular bone-derivedosteoblasts. Exp Cell Res, 2003, 291(1): 201-211

[17] Watanabe-Takano H, Takano K, Keduka E, et al. M-Ras is activated by bone morphogenetic protein-2 and participates in osteoblastic determination, differentiation, and transdifferentiation. Exp Cell Res, 2010, 316(3): 477-490

[18] Phimphilai M, Zhao Z, Boules H, et al. BMP signaling is required for RUNX2-dependent induction of the osteoblast phenotype. J Bone Miner Res, 2006, 21(4): 637-646

[19] Chang S F, Chang T K, Peng H H, et al. BMP-4 induction of arrest and differentiation of osteoblast-like cells via p21CIP1 and p27KIP1 regulation. Mol Endocrinol, 2009, 23(11): 1827-1838

[20] 赵 丹, 王 箭, 罗进勇, 等. 骨形态发生蛋白 9 通过 p38 激酶途径调控间充质干细胞 C3H10T1/2 成骨分化. 生物化学与生物物理进展, 2011, 38(11): 1001-1010 Zhao D, Wang J, Luo J Y, et al. Prog Biochem Biophys, 2011,38(11): 1001-1010

[21] Widmann C, Gibson S, Jarpe M B, et al. Mitogen-activated protein kinase: conservation of a three-kinase module from yeast to human. Physiol Rev, 1999, 79(1): 143-180

[22] Chen D, Zhao M, Mundy G R. Bone morphogenetic proteins. Growth Factors, 2004, 22(4): 233-241

[23] Xiao G, Gopalakrishnan R, Jiang D, et al. Bone morphogenetic proteins, extracellular matrix, and mitogen-activated protein kinase signaling pathways are required for osteoblast-specific gene expression and differentiation in MC3T3-E1 cells. J Bone Miner Res, 2002, 17(1): 101-110

[24] Celil A B, Campbell P G. BMP-2 and insulin-like growth factor-I mediate osterix (Osx) expression in human mesenchymal stem cells via the MAPK and protein kinase D signaling pathways. J Biol Chem, 2005, 280(36): 31353-31359

[25] Luo J, Tang M, Huang J, et al. TGF β/BMP typeⅠ receptors ALK1 and ALK2 are essential for BMP9-induced osteogenic signaling in mesenchymal stem cells. J Biol Chem, 2010, 285 (38): 29588- 29598

[26] Wu N, Zhao Y, Yin Y, et al. Identification and analysis of type Ⅱ TGF-b receptors in BMP-9-induced osteogenicdifferentiation of C3H10T1/2 mesenchymal stem cells. Acta Biochim Biophys Sin, 2010, 42(10): 699-708

[27] Guicheux J, Lemonnier J, Ghayor C, et al Activation of p38 mitogen-activated protein kinase and c-Jun-NH2-terminal kinase by BMP-2 and their implication in the stimulation of osteoblastic cell differentiation. J Bone Miner Res, 2003, 18(11): 2060-2068

[28] Gallea S, Lallemand F, Atfi A, et al. Activation of mitogenactivated protein kinase cascades is involved in regulation of bone morphogenetic protein-2-induced osteoblast differentiation in pluripotent C2C12 cells. Bone, 2001, 28(5): 491-498

[29] Huang Y F, Lin J J, Lin C H, et al. c-Jun N-terminal kinase 1 negatively regulates osteoblastic differentiation induced by BMP2 via phosphorylation of Runx2 at Ser104. J Bone Miner Res, 2012,27(5): 1093-1105