Nicotine derived genotoxic effects in human primary parotid gland cells as assessed in vitro by comet assay, cytokinesis-block micronucleus test and chromosome aberrations test.
Genotoxic effects of nicotine were described in different human cells including salivary gland cells. Based on the high nicotine concentration in saliva of smokers or patients using therapeutic nicotine patches, the current study was performed to evaluate the genotoxic potential of nicotine in human salivary gland cells. Therefore, primary salivary gland cells from 10 patients undergoing parotid gland surgery were exposed to nicotine concentrations between 1 μM and 1000 μM for 1 h in the absence of exogenous metabolic activation. The acinar phenotype was proven by immunofluorescent staining of alpha-amylase. Genotoxic effects were evaluated using the Comet assay, the micronucleus test and the chromosome aberration test. Cytotoxicity and apoptosis were determined by trypan blue exclusion test and Caspase-3 assay. Nicotine was able to induce genotoxic effects in all three assays. The chromosome aberration test was the most sensitive and increases in numerical and structural (chromatid-type and chromosome-type) aberrations were seen at ≥1 μM, whereas increases in micronuclei frequency were detected at 10 μM and DNA damage as measured in the Comet assay was noted at >100 μM. No cytotoxic damage or influence of apoptosis could be demonstrated. Nicotine as a possible risk factor for tumor initiation in salivary glands is still discussed controversially. Our results demonstrated the potential of nicotine to induce genotoxic effects in salivary gland cells. These results were observed at saliva nicotine levels similar to those found after oral or transdermal exposure to nicotine and suggest the necessity of careful monitoring of the use of nicotine in humans.
Department of Oto-Rhino-Laryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University Hospital of Wuerzburg, Josef-Schneider-Str. 11, D-97080 Wuerzburg, Germany. Electronic address: ginzkey_c@ukw.de.
Christian
C
Ginzkey
Department of Oto-Rhino-Laryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University Hospital of Wuerzburg, Josef-Schneider-Str. 11, D-97080 Wuerzburg, Germany. Electronic address: friehs_g@ukw.de.
Gudrun
G
Steussloff
Department of Oto-Rhino-Laryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University Hospital of Wuerzburg, Josef-Schneider-Str. 11, D-97080 Wuerzburg, Germany. Electronic address: koehler_c@ukw.de.
Christian
C
Koehler
Department of Oto-Rhino-Laryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University Hospital of Wuerzburg, Josef-Schneider-Str. 11, D-97080 Wuerzburg, Germany. Electronic address: burghartz_m@ukw.de.
Marc
M
Burghartz
Department of Oto-Rhino-Laryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University Hospital of Wuerzburg, Josef-Schneider-Str. 11, D-97080 Wuerzburg, Germany. Electronic address: scherzed_a@ukw.de.
Agmal
A
Scherzed
Department of Oto-Rhino-Laryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University Hospital of Wuerzburg, Josef-Schneider-Str. 11, D-97080 Wuerzburg, Germany. Electronic address: hackenberg_s@ukw.de.
Stephan
S
Hackenberg
Department of Oto-Rhino-Laryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University Hospital of Wuerzburg, Josef-Schneider-Str. 11, D-97080 Wuerzburg, Germany. Electronic address: hagen_r@ukw.de.
Rudolf
R
Hagen
Department of Oto-Rhino-Laryngology, Plastic, Aesthetic and Reconstructive Head and Neck Surgery, University Hospital of Wuerzburg, Josef-Schneider-Str. 11, D-97080 Wuerzburg, Germany. Electronic address: kleinsasser_n@ukw.de.
Norbert H
NH
Kleinsasser
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002869:Chromosome Aberrations / Q000139:chemically induced; D020552:Comet Assay; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D015162:Micronucleus Tests; D008875:Middle Aged; D009153:Mutagens / Q000633:toxicity*; D009538:Nicotine / Q000633:toxicity*; D010306:Parotid Gland / Q000166:cytology*; D000516:alpha-Amylases / Q000378:metabolism
D009153:Mutagens; D009538:Nicotine; D000516:alpha-Amylases; D053148:Caspase 3
Chromosome aberration; Comet assay; Genotoxicity; Micronucleus; Nicotine; Salivary gland