title: BMP-2 overexpression augments vascular smooth muscle cell motility by upregulating myosin Va via Erk signaling.
abstract: BACKGROUND
  The disruption of physiologic vascular smooth muscle cell (VSMC) migration initiates atherosclerosis development. The biochemical mechanisms leading to dysfunctional VSMC motility remain unknown. Recently, cytokine BMP-2 has been implicated in various vascular physiologic and pathologic processes. However, whether BMP-2 has any effect upon VSMC motility, or by what manner, has never been investigated.
  
  
  METHODS
  VSMCs were adenovirally transfected to genetically overexpress BMP-2. VSMC motility was detected by modified Boyden chamber assay, confocal time-lapse video assay, and a colony wounding assay. Gene chip array and RT-PCR were employed to identify genes potentially regulated by BMP-2. Western blot and real-time PCR detected the expression of myosin Va and the phosphorylation of extracellular signal-regulated kinases 1/2 (Erk1/2). Immunofluorescence analysis revealed myosin Va expression locale. Intracellular Ca(2+) oscillations were recorded.
  
  
  RESULTS
  VSMC migration was augmented in VSMCs overexpressing BMP-2 in a dose-dependent manner. siRNA-mediated knockdown of myosin Va inhibited VSMC motility. Both myosin Va mRNA and protein expression significantly increased after BMP-2 administration and were inhibited by Erk1/2 inhibitor U0126. BMP-2 induced Ca(2+) oscillations, generated largely by a "cytosolic oscillator".
  
  
  CONCLUSIONS
  BMP-2 significantly increased VSMCs migration and myosin Va expression, via the Erk signaling pathway and intracellular Ca(2+) oscillations. We provide additional insight into the pathophysiology of atherosclerosis, and inhibition of BMP-2-induced myosin Va expression may represent a potential therapeutic strategy.
authors.affiliation: Department of Cardiology, Beijing An Zhen Hospital, Capital Medical University, and Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing 100029, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of Cardiology, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China.
authors.forename: Min
authors.initials: M
authors.lastname: Yang
authors.affiliation: Department of Nephrology, First Hospital of Tsinghua University, Beijing 100016, China.
authors.forename: Li-ping
authors.initials: LP
authors.lastname: Liu
authors.affiliation: Department of Emergency Medicine, Thomas Jefferson University, Philadelphia, PA 19107, USA.
authors.forename: Wayne Bond
authors.identifier: 0000-0002-8064-8290
authors.initials: WB
authors.lastname: Lau
authors.affiliation: Department of Cardiology, Beijing An Zhen Hospital, Capital Medical University, and Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing 100029, China.
authors.forename: Hai
authors.initials: H
authors.lastname: Gao
authors.affiliation: Department of Cardiology, Beijing An Zhen Hospital, Capital Medical University, and Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing 100029, China.
authors.forename: Man-kun
authors.initials: MK
authors.lastname: Xin
authors.affiliation: Department of Cardiology, Beijing An Zhen Hospital, Capital Medical University, and Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing 100029, China.
authors.forename: Li-Xiao
authors.initials: LX
authors.lastname: Su
authors.affiliation: Department of Cardiology, Beijing An Zhen Hospital, Capital Medical University, and Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing 100029, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Cardiology, Beijing An Zhen Hospital, Capital Medical University, and Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing 100029, China.
authors.forename: Shu-Juan
authors.initials: SJ
authors.lastname: Cheng
authors.affiliation: Department of Cardiology, Beijing An Zhen Hospital, Capital Medical University, and Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing 100029, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Fan
authors.affiliation: Department of Cardiology, Beijing An Zhen Hospital, Capital Medical University, and Beijing Institute of Heart, Lung and Blood Vessel Disease, Beijing 100029, China.
authors.forename: Jing-Hua
authors.initials: JH
authors.lastname: Liu
mesh_terms: D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002465:Cell Movement; D002478:Cells, Cultured; D018715:Microscopy, Video; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D018995:Myosin Heavy Chains / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D024701:Myosin Type V / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D015854:Up-Regulation
chemical_list: D055396:Bone Morphogenetic Protein 2; C513081:Myo5a protein, rat; D034741:RNA, Small Interfering; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D024701:Myosin Type V; D018995:Myosin Heavy Chains; D002118:Calcium