Accelerated repair and reduced mutagenicity of DNA damage induced by cigarette smoke in human bronchial cells transfected with E.coli formamidopyrimidine DNA glycosylase.
Cigarette smoke (CS) is associated to a number of pathologies including lung cancer. Its mutagenic and carcinogenic effects are partially linked to the presence of reactive oxygen species and polycyclic aromatic hydrocarbons (PAH) inducing DNA damage. The bacterial DNA repair enzyme formamidopyrimidine DNA glycosylase (FPG) repairs both oxidized bases and different types of bulky DNA adducts. We investigated in vitro whether FPG expression may enhance DNA repair of CS-damaged DNA and counteract the mutagenic effects of CS in human lung cells. NCI-H727 non small cell lung carcinoma cells were transfected with a plasmid vector expressing FPG fused to the Enhanced Green Fluorescent Protein (EGFP). Cells expressing the fusion protein EGFP-FPG displayed accelerated repair of adducts and DNA breaks induced by CS condensate. The mutant frequencies induced by low concentrations of CS condensate to the Na(+)K(+)-ATPase locus (oua(r)) were significantly reduced in cells expressing EGFP-FPG. Hence, expression of the bacterial DNA repair protein FPG stably protects human lung cells from the mutagenic effects of CS by improving cells' capacity to repair damaged DNA.
Department of Health Sciences, University of Genova, Genova, Italy ; Mutagenesis Unit, IRCCS Azienda Ospedaliera Universitaria San Martino - IST Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Mara
M
Foresta
Department of Health Sciences, University of Genova, Genova, Italy ; Mutagenesis Unit, IRCCS Azienda Ospedaliera Universitaria San Martino - IST Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Alberto
A
Izzotti
Department of Health Sciences, University of Genova, Genova, Italy.
Sebastiano
S
La Maestra
Department of Health Sciences, University of Genova, Genova, Italy.
Rosanna
R
Micale
Molecular Oncology and Angiogenesis Unit, IRCCS Azienda Ospedaliera Universitaria San Martino - IST Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Alessandro
A
Poggi
Mutagenesis Unit, IRCCS Azienda Ospedaliera Universitaria San Martino - IST Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Donatella
D
Vecchio
Mutagenesis Unit, IRCCS Azienda Ospedaliera Universitaria San Martino - IST Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
Guido
G
Frosina
D001980:Bronchi / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D004249:DNA Damage; D004260:DNA Repair; D043382:DNA-Formamidopyrimidine Glycosylase / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics; D005260:Female; D006801:Humans; D008875:Middle Aged; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014028:Tobacco Smoke Pollution / Q000009:adverse effects*; D014162:Transfection
D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D014028:Tobacco Smoke Pollution; D043382:DNA-Formamidopyrimidine Glycosylase; C468610:DNA-formamidopyrimidine glycosylase, E coli