Leonardo Lupacchini1, Fabrizio Maggi2, Carlo Tomino3, Chiara De Dominicis1, Cristiana Mollinari4,5, Massimo Fini3, Stefano Bonassi6,7, Daniela Merlo5, Patrizia Russo6,7. 1. Molecular and Cellular Neurobiology, IRCSS San Raffaele Pisana, Via di Val Cannuta, 247, I-00166 Rome, Italy. 2. Department of Medicine and Surgery, and Laboratory of Clinical Microbiology, University of Insubria, ASST dei Sette Laghi, viale Luigi Borri 57, I-21100 Varese, Italy. 3. Scientific Direction, IRCSS San Raffaele Pisana, Via di Val Cannuta, 247, I-00166 Rome, Italy. 4. Institute of Translational Pharmacology, National Research Council, Via Fosso del Cavaliere 100, 00133 Rome, Italy. 5. Department of Neuroscience, Istituto Superiore di Sanità, Viale Regina Elena 299, I-00161 Rome, Italy. 6. Clinical and Molecular Epidemiology, IRCSS San Raffaele Pisana, Via di Val Cannuta, 247, I-00166 Rome, Italy. 7. Department of Human Sciences and Quality of Life Promotion San Raffaele University, Via di Val Cannuta, 247, I-00166 Rome, Italy.
Abstract
(1) Background: Nicotine is implicated in the SARS-COV-2 infection through activation of the α7-nAChR and over-expression of ACE2. Our objective was to clarify the role of nicotine in SARS-CoV-2 infection exploring its molecular and cellular activity. (2) Methods: HBEpC or si-mRNA-α7-HBEpC were treated for 1 h, 48 h or continuously with 10-7 M nicotine, a concentration mimicking human exposure to a cigarette. Cell viability and proliferation were evaluated by trypan blue dye exclusion and cell counting, migration by cell migration assay, senescence by SA-β-Gal activity, and anchorage-independent growth by cloning in soft agar. Expression of Ki67, p53/phospho-p53, VEGF, EGFR/pEGFR, phospho-p38, intracellular Ca2+, ATP and EMT were evaluated by ELISA and/or Western blotting. (3) Results: nicotine induced through α7-nAChR (i) increase in cell viability, (ii) cell proliferation, (iii) Ki67 over-expression, (iv) phospho-p38 up-regulation, (v) EGFR/pEGFR over-expression, (vi) increase in basal Ca2+ concentration, (vii) reduction of ATP production, (viii) decreased level of p53/phospho-p53, (ix) delayed senescence, (x) VEGF increase, (xi) EMT and consequent (xii) enhanced migration, and (xiii) ability to grow independently of the substrate. (4) Conclusions: Based on our results and on evidence showing that nicotine potentiates viral infection, it is likely that nicotine is involved in SARS-CoV-2 infection and severity.
(1) Background: Nicotine is implicated in the SARS-COV-2 infection through activation of the α7-nAChR and over-expression of ACE2. Our objective was to clarify the role of nicotine in SARS-CoV-2 infection exploring its molecular and cellular activity. (2) Methods: HBEpC or si-mRNA-α7-HBEpC were treated for 1 h, 48 h or continuously with 10-7 M nicotine, a concentration mimicking human exposure to a cigarette. Cell viability and proliferation were evaluated by trypan blue dye exclusion and cell counting, migration by cell migration assay, senescence by SA-β-Gal activity, and anchorage-independent growth by cloning in soft agar. Expression of Ki67, p53/phospho-p53, VEGF, EGFR/pEGFR, phospho-p38, intracellular Ca2+, ATP and EMT were evaluated by ELISA and/or Western blotting. (3) Results:nicotine induced through α7-nAChR (i) increase in cell viability, (ii) cell proliferation, (iii) Ki67 over-expression, (iv) phospho-p38 up-regulation, (v) EGFR/pEGFR over-expression, (vi) increase in basal Ca2+ concentration, (vii) reduction of ATP production, (viii) decreased level of p53/phospho-p53, (ix) delayed senescence, (x) VEGF increase, (xi) EMT and consequent (xii) enhanced migration, and (xiii) ability to grow independently of the substrate. (4) Conclusions: Based on our results and on evidence showing that nicotine potentiates viral infection, it is likely that nicotine is involved in SARS-CoV-2 infection and severity.
Authors: Luigi Sansone; Antonio de Iure; Mario Cristina; Manuel Belli; Laura Vitiello; Federica Marcolongo; Alfredo Rosellini; Lisa Macera; Pietro Giorgio Spezia; Carlo Tomino; Stefano Bonassi; Matteo A Russo; Fabrizio Maggi; Patrizia Russo Journal: Int J Mol Sci Date: 2022-08-22 Impact factor: 6.208
Authors: Francesca Lunardi; Francesco Fortarezza; Luca Vedovelli; Federica Pezzuto; Annalisa Boscolo; Marco Rossato; Roberto Vettor; Anna Maria Cattelan; Claudia Del Vecchio; Andrea Crisanti; Paolo Navalesi; Dario Gregori; Fiorella Calabrese Journal: Biomolecules Date: 2021-05-26