6533b873fe1ef96bd12d4e80

RESEARCH PRODUCT

Carbocysteine counteracts the effects of cigarette smoke on cell growth and on the SIRT1/FoxO3 axis in bronchial epithelial cells

S. Di VincenzoElisabetta PaceMark GjomarkajAndreina BrunoSalvatore BattagliaPaola DinoMaria R. BonsignoreMaria FerraroLuigi LanataFederico Saibene

subject

Bronchial epithelial cell0301 basic medicineSenescenceAgingPathologymedicine.medical_specialtyApoptosisSettore MED/10 - Malattie Dell'Apparato RespiratorioBiologyBiochemistryCell LineFlow cytometry03 medical and health sciencesSIRT10302 clinical medicineEndocrinologyGeneticSirtuin 1Western blotSmokeTobaccoSurvivinGeneticsmedicineHumansClonogenic assayMolecular BiologyCellular SenescenceCell ProliferationRegulation of gene expressionmedicine.diagnostic_testCell growthCarbocysteineForkhead Box Protein O3Cigarette smokeEpithelial CellsCarbocysteineCell BiologyCell biologyOxidative Stress030104 developmental biology030220 oncology & carcinogenesisFoxO3

description

Abstract Background Cigarette smoke may accelerate cellular senescence by increasing oxidative stress. Altered proliferation and altered expression of anti-aging factors, including SIRT1 and FoxO3, characterise cellular senescence. The effects of carbocysteine on the SIRT1/FoxO3 axis and on downstream molecular mechanisms in human bronchial epithelial cells exposed to cigarette smoke are largely unknown. Aims Aim of this study was to explore whether carbocysteine modulated SIRT1/FoxO3 axis, and downstream molecular mechanisms associated to cellular senescence, in a bronchial epithelial cell line (16-HBE) exposed to cigarette smoke. Methods 16HBE cells were stimulated with/without cigarette smoke extracts (CSE) and carbocysteine. Flow cytometry and clonogenic assay were used to assess cell proliferation; western blot analysis was used for assessing nuclear expression of SIRT1 and FoxO3. The nuclear co-localization of SIRT1 and FoxO3 was assessed by fluorescence microscopy. Beta galactosidase (a senescence marker) and SIRT1 activity were assessed by specific staining and colorimetric assays, respectively. ChiP Assay and flow cytometry were used for assessing survivin gene regulation and protein expression, respectively. Results CSE decreased cell proliferation, the nuclear expression of SIRT1 and FoxO3 and increased beta galactosidase staining. CSE, reduced SIRT1 activity and FoxO3 localization on survivin promoter thus increasing survivin expression. In CSE stimulated bronchial epithelial cells carbocysteine reverted these phenomena by increasing cell proliferation, and SIRT1 and FoxO3 nuclear expression, and by reducing beta galactosidase staining and survivin expression. Conclusions The study shows for the first time that carbocysteine may revert some senescence processes induced by oxidative stress due to cigarette smoke exposure.

yearjournalcountryeditionlanguage
2016-03-30Experimental Gerontology
https://doi.org/10.1016/j.exger.2016.05.013