title: NCK Associated Protein 1 Modulated by miRNA-214 Determines Vascular Smooth Muscle Cell Migration, Proliferation, and Neointima Hyperplasia.
abstract: MicroRNA miR-214 has been implicated in many biological cellular functions, but the impact of miR-214 and its target genes on vascular smooth muscle cell (VSMC) proliferation, migration, and neointima smooth muscle cell hyperplasia is unknown.
  
  
  
  Expression of miR-214 was closely regulated by different pathogenic stimuli in VSMCs through a transcriptional mechanism and decreased in response to vascular injury. Overexpression of miR-214 in serum-starved VSMCs significantly decreased VSMC proliferation and migration, whereas knockdown of miR-214 dramatically increased VSMC proliferation and migration. Gene and protein biochemical assays, including proteomic analyses, showed that NCK associated protein 1 (NCKAP1)-a major component of the WAVE complex that regulates lamellipodia formation and cell motility-was negatively regulated by miR-214 in VSMCs. Luciferase assays showed that miR-214 substantially repressed wild-type but not the miR-214 binding site mutated version of NCKAP1 3' untranslated region luciferase activity in VSMCs. This result confirmed that NCKAP1 is the functional target of miR-214 in VSMCs. NCKAP1 knockdown in VSMCs recapitulates the inhibitory effects of miR-214 overexpression on actin polymerization, cell migration, and proliferation. Data from cotransfection experiments also revealed that inhibition of NCKAP1 is required for miR-214-mediated lamellipodia formation, cell motility, and growth. Importantly, locally enforced expression of miR-214 in the injured vessels significantly reduced NCKAP1 expression levels, inhibited VSMC proliferation, and prevented neointima smooth muscle cell hyperplasia after injury.
  
  
  
  We uncovered an important role of miR-214 and its target gene NCKAP1 in modulating VSMC functions and neointima hyperplasia. Our findings suggest that miR-214 represents a potential therapeutic target for vascular diseases.
authors.affiliation: Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom.
authors.forename: Tayyab Adeel
authors.initials: TA
authors.lastname: Afzal
authors.affiliation: Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom.
authors.forename: Le Anh
authors.initials: LA
authors.lastname: Luong
authors.affiliation: Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom.|Department of Cardiothoracic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Chen
authors.affiliation: Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom.|Department of Cardiothoracic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom.|Department of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Yang
authors.affiliation: Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom.|Department of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Qishan
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom.
authors.forename: Weiwei
authors.initials: W
authors.lastname: An
authors.affiliation: Centre for Haemato-Oncology, Barts Cancer Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom.
authors.forename: Edmund
authors.initials: E
authors.lastname: Wilkes
authors.affiliation: Translational Medicine & Therapeutics, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom.
authors.forename: Kenta
authors.initials: K
authors.lastname: Yashiro
authors.affiliation: Centre for Haemato-Oncology, Barts Cancer Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom.
authors.forename: Pedro R
authors.initials: PR
authors.lastname: Cutillas
authors.affiliation: Department of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China q.xiao@qmul.ac.uk li.zhang.uk@googlemail.com.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Centre for Clinical Pharmacology, William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, United Kingdom q.xiao@qmul.ac.uk li.zhang.uk@googlemail.com.
authors.forename: Qingzhong
authors.initials: Q
authors.lastname: Xiao
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000494:pharmacology; D000818:Animals; D000077214:Becaplermin; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology; D015536:Down-Regulation; D005263:Femoral Artery / Q000601:surgery; D055785:Gene Knockdown Techniques; D006965:Hyperplasia / Q000473:pathology; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000378:metabolism / Q000502:physiology*; D009131:Muscle, Smooth, Vascular / Q000502:physiology; D009154:Mutation / Q000235:genetics; D032389:Myocytes, Smooth Muscle; D058426:Neointima / Q000473:pathology*; D040901:Proteomics; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000502:physiology; D051879:Twist-Related Protein 1 / Q000037:antagonists & inhibitors
chemical_list: D043925:Angiogenesis Inducing Agents; D008565:Membrane Proteins; D035683:MicroRNAs; C534781:Mirn214 microRNA, mouse; C502089:Nckap1 protein, mouse; D020574:Proto-Oncogene Proteins c-sis; D034741:RNA, Small Interfering; D051879:Twist-Related Protein 1; D000077214:Becaplermin
keywords: NCK‐associated protein 1; cell migration; cell proliferation; miRNA‐214; microRNA; neointimal hyperplasia; vascular biology; vascular disease; vascular smooth muscle