Login Register Cart 0
Generic placeholder image

Current Molecular Medicine

Editor-in-Chief

ISSN (Print): 1566-5240
ISSN (Online): 1875-5666

Back
Journal Home About Journal Editorial Board Journal Insight Current Issue Volumes/Issues
Subscribe Translate in Chinese
Research Article

Cytochalasin D Promotes Osteogenic Differentiation of MC3T3-E1 Cells via p38-MAPK Signaling Pathway

Author(s): Qingcheng Liu, Yu Zhuang, Ningjuan Ouyang* and Hongbo Yu*

Volume 20, Issue 1, 2020

Page: [79 - 88] Pages: 10

DOI: 10.2174/1566524019666191007104816

open access plus

Abstract

Background: Bone defect caused by trauma, tumor resection, infection or congenital malformation is a common clinical disease. Bone tissue engineering is regarded as a promising way of bone defect reconstruction. Thus, agents that can promote osteogenesis have received great attention. Cytochalasin D (Cyto D), a metabolite derived from molds, proves to be able to modify actin, reorganize cytoskeleton, and then promote the osteogenic differentiation.

Objective: The purpose of this study was to explore the effect and mechanism of Cyto D on osteogenic differentiation of mouse pre-osteoblast MC3T3-E1 cells.

Methods: The optimum concentration of Cyto D was explored. The osteogenic differentiation of MC3T3-E1 cells induced by Cyto D was assessed by alkaline phosphatase (ALP) staining, Alizarin Red S (ARS) staining, western blotting and quantitative real-time polymerase chain reaction (RT-qPCR). In addition, a specific pathway inhibitor was utilized to explore whether MAPK pathways were involved in this process.

Results: The results showed that the optimized concentration of action was 10-2µg/ml. The expression of Runx2, OCN and OSX was up-regulated by the supplement of Cyto D. ALP activity, calcium deposition, and phosphorylation level of p38 protein were also improved. Inhibition of the pathway significantly reduced the activation of p38, and the expression of osteogenic-related genes.

Conclusion: Cyto D can promote the osteogenic differentiation of MC3T3 cells via the p38-MAPK signaling pathway, but not the ERK1/2 or JNK, and it is a potential agent to improve the osteogenesis of MC3T3 cells.

Keywords: Bone remodeling, cytochalasin D, osteogenic differentiation, MC3T3-E1 cells, MAPK, p38.

« Previous
[1]
Fernandez-Yague MA, Abbah SA, McNamara L, Zeugolis DI, Pandit A, Biggs MJ. Biomimetic approaches in bone tissue engineering: Integrating biological and physicomechanical strategies. Adv Drug Deliv Rev 2015; 84: 1-29.
[http://dx.doi.org/10.1016/j.addr.2014.09.005] [PMID: 25236302]
[2]
Wang Z, Wu D, Zou J, et al. Development of demineralized bone matrix based implantable and biomimetic microcarrier for stem cell expansion and single-step tissue-engineered bone graft construction. J Mater Chem B Mater Biol Med 2016; 5(1)
[http://dx.doi.org/10.1039/C6TB02414A]
[3]
Li K, Zhang S, Wang S, et al. Positive effect of magnetic-conductive bifunctional fibrous scaffolds on guiding double electrical and magnetic stimulations to pre-osteoblasts. J Biomed Nanotechnol 2019; 15(3): 477-86.
[http://dx.doi.org/10.1166/jbn.2019.2708] [PMID: 31165693]
[4]
Dudakovic A, Camilleri ET, Riester SM, et al. Enhancer of Zeste Homolog 2 inhibition stimulates bone formation and mitigates bone loss caused by ovariectomy in skeletally mature mice. J Biol Chem 2016; 291(47): 24594-606.
[http://dx.doi.org/10.1074/jbc.M116.740571] [PMID: 27758858]
[5]
Hemming S, Cakouros D, Isenmann S, et al. EZH2 and KDM6A act as an epigenetic switch to regulate mesenchymal stem cell lineage specification. Stem Cells 2014; 32(3): 802-15.
[http://dx.doi.org/10.1002/stem.1573] [PMID: 24123378]
[6]
Hemming S, Cakouros D, Codrington J, et al. EZH2 deletion in early mesenchyme compromises postnatal bone microarchitecture and structural integrity and accelerates remodeling. FASEB J 2017; 31(3): 1011-27.
[http://dx.doi.org/10.1096/fj.201600748R] [PMID: 27934660]
[7]
Sen B, Xie Z, Uzer G, et al. Intranuclear actin regulates osteogenesis. Stem Cells 2015; 33(10): 3065-76.
[http://dx.doi.org/10.1002/stem.2090] [PMID: 26140478]
[8]
Samsonraj RM, Dudakovic A, Manzar B, et al. Osteogenic stimulation of human adipose-derived mesenchymal stem cells using a fungal metabolite that suppresses the polycomb group protein EZH2. Stem Cells Transl Med 2018; 7(2): 197-209.
[http://dx.doi.org/10.1002/sctm.17-0086] [PMID: 29280310]
[9]
Sen B, Uzer G, Samsonraj RM, et al. Intranuclear Actin Structure Modulates Mesenchymal Stem Cell Differentiation. Stem Cells 2017; 35(6): 1624-35.
[http://dx.doi.org/10.1002/stem.2617] [PMID: 28371128]
[10]
Engler AJ, Sen S, Sweeney HL, Discher DE. Matrix elasticity directs stem cell lineage specification. Cell 2006; 126(4): 677-89.
[http://dx.doi.org/10.1016/j.cell.2006.06.044] [PMID: 16923388]
[11]
McBeath R, Pirone DM, Nelson CM, Bhadriraju K, Chen CS. Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell 2004; 6(4): 483-95.
[http://dx.doi.org/10.1016/S1534-5807(04)00075-9] [PMID: 15068789]
[12]
Le HQ, Ghatak S, Yeung CYC, et al. Mechanical regulation of transcription controls Polycomb-mediated gene silencing during lineage commitment. Nat Cell Biol 2016; 18(8): 864-75.
[http://dx.doi.org/10.1038/ncb3387] [PMID: 27398909]
[13]
Heo SJ, Thorpe SD, Driscoll TP, Duncan RL, Lee DA, Mauck RL. Biophysical regulation of chromatin architecture instills a mechanical memory in mesenchymal stem cells. Sci Rep 2015; 5: 16895.
[http://dx.doi.org/10.1038/srep16895] [PMID: 26592929]
[14]
Dopie J, Skarp K-P, Rajakylä EK, Tanhuanpää K, Vartiainen MK. Active maintenance of nuclear actin by importin 9 supports transcription. Proc Natl Acad Sci USA 2012; 109(9): E544-52.
[http://dx.doi.org/10.1073/pnas.1118880109] [PMID: 22323606]
[15]
Samsonraj RM, Paradise CR, Dudakovic A, et al. Validation of osteogenic properties of cytochalasin d by high-resolution rna-sequencing in mesenchymal stem cells derived from bone marrow and adipose tissues. Stem Cells Dev 2018; 27(16): 1136-45.
[http://dx.doi.org/10.1089/scd.2018.0037] [PMID: 29882479]
[16]
Majidinia M, Sadeghpour A, Yousefi B. The roles of signaling pathways in bone repair and regeneration. J Cell Physiol 2018; 233(4): 2937-48.
[http://dx.doi.org/10.1002/jcp.26042] [PMID: 28590066]
[17]
Johnson GL, Lapadat R. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science 2002; 298(5600): 1911-2.
[http://dx.doi.org/10.1126/science.1072682] [PMID: 12471242]
[18]
Cong Q, Jia H, Li P, et al. p38α MAPK regulates proliferation and differentiation of osteoclast progenitors and bone remodeling in an aging-dependent manner. Sci Rep 2017; 7: 45964.
[http://dx.doi.org/10.1038/srep45964] [PMID: 28382965]
[19]
Jessop HL, Rawlinson SCF, Pitsillides AA, Lanyon LE. Mechanical strain and fluid movement both activate extracellular regulated kinase (ERK) in osteoblast-like cells but via different signaling pathways. Bone 2002; 31(1): 186-94.
[http://dx.doi.org/10.1016/S8756-3282(02)00797-4] [PMID: 12110433]
[20]
Matsuda N, Morita N, Matsuda K, Watanabe M. Proliferation and differentiation of human osteoblastic cells associated with differential activation of MAP kinases in response to epidermal growth factor, hypoxia, and mechanical stress in vitro. Biochem Biophys Res Commun 1998; 249(2): 350-4.
[http://dx.doi.org/10.1006/bbrc.1998.9151] [PMID: 9712699]
[21]
Wang T, Zhai Y, Nuzzo M, Yang X, Yang Y, Zhang X. Layer-by-layer nanofiber-enabled engineering of biomimetic periosteum for bone repair and reconstruction. Biomaterials 2018; 182: 279-88.
[http://dx.doi.org/10.1016/j.biomaterials.2018.08.028] [PMID: 30142527]
[22]
Fan Z, Yamaza T, Lee JS, et al. BCOR regulates mesenchymal stem cell function by epigenetic mechanisms. Nat Cell Biol 2009; 11(8): 1002-9.
[http://dx.doi.org/10.1038/ncb1913] [PMID: 19578371]
[23]
Yu J, He H, Tang C, et al. Differentiation potential of STRO-1+ dental pulp stem cells changes during cell passaging. BMC Cell Biol 2010; 11(1): 32.
[http://dx.doi.org/10.1186/1471-2121-11-32] [PMID: 20459680]
[24]
Kustermans G, Piette J, Legrand-Poels S. Actin-targeting natural compounds as tools to study the role of actin cytoskeleton in signal transduction. Biochem Pharmacol 2008; 76(11): 1310-22.
[http://dx.doi.org/10.1016/j.bcp.2008.05.028] [PMID: 18602087]
[25]
Komori T. Regulation of osteoblast differentiation by transcription factors. J Cell Biochem 2006; 99(5): 1233-9.
[http://dx.doi.org/10.1002/jcb.20958] [PMID: 16795049]
[26]
Harada S, Rodan GA. Control of osteoblast function and regulation of bone mass. Nature 2003; 423(6937): 349-55.
[http://dx.doi.org/10.1038/nature01660] [PMID: 12748654]
[27]
Termaat MF, Den Boer FC, Bakker FC, Patka P, Haarman HJ. Bone morphogenetic proteins. Development and clinical efficacy in the treatment of fractures and bone defects. J Bone Joint Surg Am 2005; 87(6): 1367-78.
[http://dx.doi.org/10.2106/JBJS.D.02585] [PMID: 15930551]
[28]
Lemonnier J, Ghayor C, Guicheux J, Caverzasio J. Protein kinase C-independent activation of protein kinase D is involved in BMP-2-induced activation of stress mitogen-activated protein kinases JNK and p38 and osteoblastic cell differentiation. J Biol Chem 2004; 279(1): 259-64.
[http://dx.doi.org/10.1074/jbc.M308665200] [PMID: 14573624]
[29]
Zhang P, Dai Q, Ouyang N, et al. Mechanical strain promotes osteogenesis of bmscs from ovariectomized rats via the erk1/2 but not p38 or jnk-mapk signaling pathways. Curr Mol Med 2015; 15(8): 780-9.
[http://dx.doi.org/10.2174/1566524015666150824143830] [PMID: 26299771]
[30]
Zhang W, Guo H, Jing H, et al. Lactoferrin stimulates osteoblast differentiation through PKA and p38 pathways independent of lactoferrin’s receptor LRP1. J Bone Miner Res 2014; 29(5): 1232-43.
[http://dx.doi.org/10.1002/jbmr.2116] [PMID: 24877241]
[31]
Wanachewin O, Boonmaleerat K, Pothacharoen P, Reutrakul V, Kongtawelert P. Sesamin stimulates osteoblast differentiation through p38 and ERK1/2 MAPK signaling pathways. BMC Complement Altern Med 2012; 12: 71.
[http://dx.doi.org/10.1186/1472-6882-12-71] [PMID: 22646286]
[32]
Guo C, Yang XG, Wang F, Ma XY. IL-1α induces apoptosis and inhibits the osteoblast differentiation of MC3T3-E1 cells through the JNK and p38 MAPK pathways. Int J Mol Med 2016; 38(1): 319-27.
[http://dx.doi.org/10.3892/ijmm.2016.2606] [PMID: 27220839]
[33]
Ge C, Xiao G, Jiang D, Franceschi RT. Critical role of the extracellular signal-regulated kinase-MAPK pathway in osteoblast differentiation and skeletal development. J Cell Biol 2007; 176(5): 709-18.
[http://dx.doi.org/10.1083/jcb.200610046] [PMID: 17325210]
[34]
Chen X, Zhang S, Chen X, et al. Emodin promotes the osteogenesis of MC3T3-E1 cells via BMP-9/Smad pathway and exerts a preventive effect in ovariectomized rats. Acta Biochim Biophys Sin (Shanghai) 2017; 49(10): 867-78.
[http://dx.doi.org/10.1093/abbs/gmx087] [PMID: 28981600]
[35]
Nöth U, Tuli R, Seghatoleslami R, et al. Activation of p38 and Smads mediates BMP-2 effects on human trabecular bone-derived osteoblasts. Exp Cell Res 2003; 291(1): 201-11.
[http://dx.doi.org/10.1016/S0014-4827(03)00386-0] [PMID: 14597420]
[36]
Otto F, Thornell AP, Crompton T, et al. Cbfa1, a candidate gene for cleidocranial dysplasia syndrome, is essential for osteoblast differentiation and bone development. Cell 1997; 89(5): 765-71.
[http://dx.doi.org/10.1016/S0092-8674(00)80259-7] [PMID: 9182764]
[37]
Nakashima K, de Crombrugghe B. Transcriptional mechanisms in osteoblast differentiation and bone formation. Trends Genet 2003; 19(8): 458-66.
[http://dx.doi.org/10.1016/S0168-9525(03)00176-8] [PMID: 12902164]
[38]
Franceschi RT, Xiao G, Jiang D, Gopalakrishnan R, Yang S, Reith E. Multiple signaling pathways converge on the Cbfa1/Runx2 transcription factor to regulate osteoblast differentiation. Connect Tissue Res 2003; 44(Suppl. 1): 109-16.
[http://dx.doi.org/10.1080/03008200390152188] [PMID: 12952183]
[39]
Greenblatt MB, Shim JH, Zou W, et al. The p38 MAPK pathway is essential for skeletogenesis and bone homeostasis in mice. J Clin Invest 2010; 120(7): 2457-73.
[http://dx.doi.org/10.1172/JCI42285] [PMID: 20551513]
[40]
Huang FY, Mei WL, Li YN, et al. The antitumour activities induced by pegylated liposomal cytochalasin D in murine models. Eur J Cancer 2012; 48(14): 2260-9.
[http://dx.doi.org/10.1016/j.ejca.2011.12.018] [PMID: 22257793]
[41]
Chen L, Shi K, Frary CE, et al. Inhibiting actin depolymerization enhances osteoblast differentiation and bone formation in human stromal stem cells. Stem Cell Res (Amst) 2015; 15(2): 281-9.
[http://dx.doi.org/10.1016/j.scr.2015.06.009] [PMID: 26209815]

Rights & Permissions Print Cite
Article Metrics
57
17
© 2024 Bentham Science Publishers | Privacy Policy