[The single cell microgelelectrophoresis technique in ecogenotoxicology].
BACKGROUND
The development of carcinoma in the upper aerodigestive tract is often associated with exposure to xenobiotics. Therefore, the identification of such tumor initiating substances is relevant. Most genotoxicity test systems require mammalian cells, human lymphocytes or cell cultures to detect genotoxicity caused by carcinogens. The single cell microgelelectrophoresis technique (Comet assay) is presented, being a sensitive method, identifying DNA strand breaks, alkali labile sites and DNA repair in human epithelial cells of the upper aerodigestive tract. It is compared to other common techniques for the identification of genotoxic damage. Future applications and contributions of the method are introduced.
Using the alkaline microgel electrophoresis assay, freshly isolated single epithelial cells are incubated with xenobiotics causing DNA strand breaks and alkali labile sites. Data are examined using a digital computer analysis. The method is described for the application of epithelial cells of the upper aerodigestive tract and compared to other procedures for the monitoring of genotoxicity. These are the Ames test identifying mutagenicity in bacteria, the sister chromatid exchange and the micronucleus test demonstrating genomic instability in lymphocytes and cultured mammalian cells.


CONCLUSIONS
The microgel electrophoresis technique is a sensitive method to detect genotoxic effects and DNA repair in human epithelia of the upper aerodigestive tract. The assay offers considerable advantages to other common genotoxicity tests. However, combining of the Comet assay with mini organ cultures allows to use repetitive incubations with xenobiotics. Furthermore, signalling selected chromosomal material by the combination of the assay with the fluorescence in situ hybridisation, DNA-damage and -repair mechanisms within comets can be identified.
Harréus
U A
UA
Klinisch experimentelle Onkologie, Klinik und Poliklinik für Hals-, Nasen- und Ohrenkranke Ludwig-Maximilians-Universität München. uharreus@hno.med.uni-muenchen.de
Kastenbauer
E R
ER
Wallner
B C
BC
Kleinsasser
N H
NH
D001706:Biopsy; D002869:Chromosome Aberrations; D020552:Comet Assay; D004249:DNA Damage / Q000235:genetics*; D004260:DNA Repair / Q000187:drug effects* / Q000235:genetics; D004847:Epithelial Cells / Q000187:drug effects / Q000473:pathology; D006801:Humans; D015162:Micronucleus Tests; D009152:Mutagenicity Tests; D010039:Otorhinolaryngologic Neoplasms / Q000139:chemically induced* / Q000235:genetics / Q000473:pathology; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000473:pathology; D012680:Sensitivity and Specificity; D012854:Sister Chromatid Exchange; D015262:Xenobiotics / Q000633:toxicity*
D015262:Xenobiotics