title: β-arrestin-1 mediates nicotine-induced metastasis through E2F1 target genes that modulate epithelial-mesenchymal transition.
abstract: Cigarette smoking is a major risk factor in the development of non-small cell lung cancer (NSCLC), which accounts for 80% of all lung cancers. Nicotine, the major addictive component of tobacco smoke, can induce proliferation, invasion, and epithelial-to-mesenchymal transition (EMT) in NSCLC cell lines and promote metastasis of NSCLC in mice. Here, we demonstrate that the scaffolding protein β-arrestin-1 is necessary for nicotine-mediated induction of mesenchymal genes vimentin and fibronectin as well as EMT regulators ZEB1 and ZEB2. Nicotine induced changes in cell morphology and ablate tight junctions consistent with EMT; β-arrestin-1, but not β-arrestin-2, was required for these changes. β-Arrestin-1 promoted the expression of the mesenchymal genes, as well as ZEB1 and ZEB2, through the mediation of the E2F1 transcription factor; this required Src kinase activity. Stimulation of multiple NSCLC cell lines with nicotine led to enhanced recruitment of β-arrestin-1 and E2F1 on vimentin, fibronectin, and ZEB1 and ZEB2 promoters. Furthermore, there was significantly more β-arrestin-1 and E2F1 associated with these promoters in human NSCLC tumors, and β-arrestin-1 levels correlated with vimentin and fibronectin levels in human NSCLC samples. A549-luciferase cells lacking β-arrestin-1 showed a significantly reduced capacity for tumor growth and metastasis when orthotopically implanted into the lungs of SCID-beige mice. Taken together, these studies reveal a novel role for β-arrestin-1 in the growth and metastasis of NSCLC.
authors.affiliation: Department of Tumor Biology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida.
authors.forename: Smitha
authors.initials: S
authors.lastname: Pillai
authors.affiliation: Department of Surgery, University of Florida, Gainesville, Florida.
authors.forename: Jose
authors.initials: J
authors.lastname: Trevino
authors.affiliation: Mayo Clinic, Jacksonville, Florida.
authors.forename: Bhupendra
authors.initials: B
authors.lastname: Rawal
authors.affiliation: National Institute of Biomedical Genomics, Kalyani, West Bengal, India.
authors.forename: Sandeep
authors.initials: S
authors.lastname: Singh
authors.affiliation: Department of Tumor Biology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida.
authors.forename: Michelle
authors.initials: M
authors.lastname: Kovacs
authors.affiliation: Department of Thoracic Oncology, Moffitt Cancer Center, Tampa, Florida.
authors.forename: Xueli
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Biostatistics, Moffitt Cancer Center, Tampla, Florida.
authors.forename: Michael
authors.initials: M
authors.lastname: Schell
authors.affiliation: Department of Thoracic Oncology, Moffitt Cancer Center, Tampa, Florida.
authors.forename: Eric
authors.initials: E
authors.lastname: Haura
authors.affiliation: Barbara Ann Karmanos Cancer Institute, Detroit, Michigan.
authors.forename: Gerold
authors.initials: G
authors.lastname: Bepler
authors.affiliation: Department of Tumor Biology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida. Srikumar.Chellappan@moffitt.org.
authors.forename: Srikumar
authors.initials: S
authors.lastname: Chellappan
mesh_terms: D000818:Animals; D019390:Arrestins / Q000502:physiology*; D045744:Cell Line, Tumor; D050687:E2F1 Transcription Factor / Q000502:physiology*; D058750:Epithelial-Mesenchymal Transition; D005353:Fibronectins / Q000032:analysis; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology; D051379:Mice; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D009538:Nicotine / Q000494:pharmacology*; D011978:Receptors, Nicotinic / Q000502:physiology; D016160:Retinoblastoma Protein / Q000502:physiology; D015398:Signal Transduction; D019108:Tight Junctions / Q000187:drug effects; D014157:Transcription Factors / Q000235:genetics; D014746:Vimentin / Q000032:analysis; D000071799:Zinc Finger E-box-Binding Homeobox 1; D000071559:beta-Arrestin 1; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins
chemical_list: C000605253:ARRB1 protein, human; C000605254:ARRB2 protein, human; C000605255:Arrb1 protein, mouse; C000605258:Arrb2 protein, mouse; D019390:Arrestins; D050687:E2F1 Transcription Factor; C493753:E2F1 protein, human; D005353:Fibronectins; D018398:Homeodomain Proteins; D011978:Receptors, Nicotinic; D016160:Retinoblastoma Protein; D014157:Transcription Factors; D014746:Vimentin; C486307:ZEB1 protein, human; D000071799:Zinc Finger E-box-Binding Homeobox 1; D000071559:beta-Arrestin 1; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins; D009538:Nicotine