Validation of single cell gel assay in human leukocytes with 18 reference compounds.
To validate the alkaline single cell gel (SCG) assay as a tool for the detection of DNA damage in human leukocytes, we investigated the in vitro activity of 18 chemicals. Thirteen of these chemicals (pyrene (PY), benzo(a)pyrene (BaP), cyclophosphamide (CP), 4-nitroquinoline-1-oxide (4NQO), bleomycin (BLM), methylmercury chloride (MMC), mitomycin C (MTC), hydrogen peroxide (HP), diepoxybutane (DEB), glutaraldehyde (GA), formaldehyde (FA), griseofulvin (GF), sodium azide (NA)) are genotoxic in at least one cell system, while five compounds (ascorbic acid (AA), glucose (GL), D-mannitol (MAN), O-vanillin (VAN), chlorophyllin (CHL)) are classified as non-genotoxic. In this in vitro SCG assay, PY, BaP and CP were positive with exogeneous metabolic activation (rat S9 mix) while 4NQO, BLM, MMC, MTC, hydrogen peroxide, and diepoxbutane were positive in the absence of metabolic activation. CHL and VAN were unexpectedly found to induce a dose-dependent increase in DNA migration. AA, GL, and MAN were negative in a non-toxic range of doses. GF gave equivocal results, while FA and GA increased DNA migration at low doses and decreased DNA migration at higher doses. This behaviour is consistent with the known DNA damaging and crosslinking properties of these compounds. These data support the sensitivity and specificity of this assay for identifying genotoxic agents.
Frenzilli
G
G
Dipartimento di Scienze dell'Uomo e dell'Ambiente, Università di Pisa, Italy. r.barale@geog.unipi.it
Bosco
E
E
Barale
R
R
D015112:4-Nitroquinoline-1-oxide / Q000633:toxicity; D000328:Adult; D000818:Animals; D001205:Ascorbic Acid / Q000633:toxicity; D001547:Benzaldehydes / Q000633:toxicity; D001564:Benzo(a)pyrene / Q000633:toxicity; D001711:Biotransformation; D001761:Bleomycin / Q000633:toxicity; D002735:Chlorophyllides / Q000633:toxicity; D003520:Cyclophosphamide / Q000633:toxicity; D004249:DNA Damage; D004852:Epoxy Compounds / Q000633:toxicity; D005260:Female; D005557:Formaldehyde / Q000633:toxicity; D005947:Glucose / Q000633:toxicity; D005976:Glutaral / Q000633:toxicity; D006118:Griseofulvin / Q000633:toxicity; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D006863:Hydrogen-Ion Concentration; D007962:Leukocytes / Q000737:chemistry*; D008353:Mannitol / Q000633:toxicity; D008767:Methylmercury Compounds / Q000633:toxicity; D008862:Microsomes, Liver / Q000378:metabolism; D016685:Mitomycin / Q000633:toxicity; D009152:Mutagenicity Tests; D011721:Pyrenes / Q000633:toxicity; D051381:Rats; D015203:Reproducibility of Results; D019810:Sodium Azide / Q000633:toxicity
D001547:Benzaldehydes; D002735:Chlorophyllides; D004852:Epoxy Compounds; D008767:Methylmercury Compounds; D011721:Pyrenes; C060843:2-vanillin; D001761:Bleomycin; D005557:Formaldehyde; D006118:Griseofulvin; D001564:Benzo(a)pyrene; D008353:Mannitol; D016685:Mitomycin; D015112:4-Nitroquinoline-1-oxide; C007366:diepoxybutane; D003520:Cyclophosphamide; D019810:Sodium Azide; D006861:Hydrogen Peroxide; C007020:chlorophyllin; D005947:Glucose; D001205:Ascorbic Acid; C004925:methylmercuric chloride; D005976:Glutaral