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Construction of a eukaryotic expression vector pEGFP-C1-BMP-2 and its effect on cell migration 被引量:1

Construction of a eukaryotic expression vector pEGFP-C1-BMP-2 and its effect on cell migration
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摘要 Objective: Bone morphogenetic proteins (BMPs) are known to play an important role in bone and cartilage development. Recent research has shown that BMPs may induce tumorigenesis and promote tumor to spread, but the molecular mechanisms have not been elucidated. The aim of the present study was to investigate the regulatory function of BMP-2 in the migration of COS-7 cells and the underlying mechanisms. Methods: Human BMP-2 genetic fragment was amplified and introduced into the pEGFP-C1 vector. After being confirmed by XhoI and BamHI digestion analyses and DNA sequencing, the recombinant pEGFP-C1-BMP-2 plasmid was transfected into COS-7 cells. The influence of BMP-2 on cell migration and cofilin activity was detected by cell scratch assay and Western blotting. Results: The recombinant pEGFP-C1-BMP-2 was effectively expressed in COS-7 cells. An increased phosphorylation of both LIMK1 and cofilin and an enhancement of cell migration were observed in cells with overexpression of BMP-2. Conclusions: A recombinant pEGFP-C1-BMP-2 vector was successfully constructed and overexpression of BMP-2 regulated the activities of the downstream molecules of the Rho GTPase signaling pathway, which might contribute to the enhancement of cell migration. Objective: Bone morphogenetic proteins (BMPs) are known to play an important role in bone and cartilage development. Recent research has shown that BMPs may induce tumorigenesis and promote tumor to spread, but the molecular mechanisms have not been elucidated. The aim of the present study was to investigate the regulatory function of BMP-2 in the migration of COS-7 cells and the underlying mechanisms. Methods: Human BMP-2 genetic fragment was amplified and introduced into the pEGFP-C1 vector. After being confirmed by XhoI and BamHI digestion analyses and DNA sequencing, the recombinant pEGFP-C1-BMP-2 plasmid was transfected into COS-7 cells. The influence of BMP-2 on cell migration and cofilin activity was detected by cell scratch assay and Western blotting. Results: The recombinant pEGFP-C1-BMP-2 was effectively expressed in COS-7 cells. An increased phosphorylation of both LIMK1 and cofilin and an enhancement of cell migration were observed in cells with overexpression of BMP-2. Conclusions: A recombinant pEGFP-C1-BMP-2 vector was successfully constructed and overexpression of BMP-2 regulated the activities of the downstream molecules of the Rho GTPase signaling pathway, which might contribute to the enhancement of cell migration.
出处 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2012年第5期356-363,共8页 浙江大学学报(英文版)B辑(生物医学与生物技术)
基金 Project supported by the National Natural Science Foundation of China(No.81000535) the Zhejiang Provincial Natural Science Foundation of China(No.Y2100508)
关键词 Cell migration Bone morphogenetic protein-2 (BMP-2) COS-7 COFILIN Cell migration, Bone morphogenetic protein-2 (BMP-2), COS-7, Cofilin
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  • 1Arnold,S.F.,Tims,E.,McGrath,B.E.,1999.Identification of bone morphogenetic proteins and their receptors in human breast cancer cell lines: importance of BMP2.Cytokine,11(12):1031-1037.[doi:10.1006/cyto.1999.0508].
  • 2Blanco Calvo,M.,Bolos Fernandez,V.,Medina Villaamil,V.,Aparicio Gallego,G.,Diaz Prado,S.,Grande Pulido,E.,2009.Biology of BMP signalling and cancer.Clin.Transl.Oncol.,11(3):126-137.[doi:10.1007/S12094-009-0328-8].
  • 3Bobinac,D.,Maric,I.,Zoricic,S.,Spanjol,J.,Dordevic,G.,Mustac,E.,Fuckar,Z.,2005.Expression of bone morphogenetic proteins in human metastatic prostate and breast cancer.Croat.Med.J.,46(3):389-396..
  • 4Fiedler,J.,Roderer,G.,Gunther,K.P.,Brenner,R.E.,2002.BMP-2,BMP-4,and PDGF-bb stimulate chemotactic migration of primary human mesenchymal progenitor cells.J. Cell Biochem.,87(3):305-312.[doi:10.1002/ jcb.10309].
  • 5Foletta,V.C.,Lim,M.A.,Soosairajah,J.,Kelly,A.P.,Stanley,E.G.,Shannon,M.,He,W.,Das,S.,Massague,J.,Bernard,O.,2003.Direct signaling by the BMP type II receptor via the cytoskeletal regulator LIMK1.J. Cell Biol.,162(6):1089-1098.[doi:10.1083/jcb.200212060].
  • 6Fong,Y.C.,Li,T.M.,Wu,C.M.,Hsu,S.F.,Kao,S.T.,Chen,R.J.,Lin,C.C.,Liu,S.C.,Wu,C.L.,Tang,C.H.,2008.BMP-2 increases migration of human chondrosarcoma cells via PI3K/Akt pathway.J. Cell.Physiol.,217(3): 846-855.[doi:10.1002/jcp.21568].
  • 7Gamell,C.,Osses,N.,Bartrons,R.,Ruckle,T.,Camps,M.,Rosa,J.L.,Ventura,F.,2008.BMP2 induction of actin cytoskeleton reorganization and cell migration requires PI3-kinase and Cdc42 activity.J. Cell.Sci.,121(Pt 23): 3960-3970.[doi:10.1242/jcs.031286].
  • 8Gamell,C.,Susperregui,A.G.,Bernard,O.,Rosa,J.L.,Ventura,F.,2011.The p38/MK2/Hsp25 pathway is required for BMP-2-induced cell migration.PLoS One,6(1):e16477.[doi:10.1371/journal.pone.0016477].
  • 9Goldstein,A.M.,Brewer,K.C.,Doyle,A.M.,Nagy,N.,Roberts,D.J.,2005.BMP signaling is necessary for neural crest cell migration and ganglion formation in the enteric nervous system.Mech.Dev.,122(6):821-833.[doi:10.1016/j.mod.2005.03.003].
  • 10Hocking,J.C.,Hehr,C.L.,Bertolesi,G.,Funakoshi,H.,Nakamura,T.,McFarlane,S.,2009.LIMK1 acts downstream of BMP signaling in developing retinal ganglion cell axons but not dendrites.Dev.Biol.,330(2): 273-285.[doi:10.1016/j.ydbio.2009.03.027].

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