Abstract
There is considerable discussion within the scientific community as to the appropriate measures of cytotoxicity to use when deciding on the maximum concentration of a substance to test in vitro for its ability to induce DNA damage using the Comet assay. Conventional cytotoxicity assessment methods, such as trypan blue dye exclusion or relative cell number (cell counts) may not be the most biologically relevant measurement for cytotoxicity in this assay. Thus, we evaluated for decreased levels of adenosine triphosphate (ATP) and activation of Caspase-3/7 as well as relative cell number and trypan blue exclusion in order to understand the correlation among test compound concentration, cytotoxicity and genotoxicity outcomes in the Comet assay. We tested two non-genotoxic and non-cytotoxic compounds (d-glucose and ethanol), two non-genotoxic but cytotoxic compounds (2,4-dichlorophenol and tunicamycin) and four genotoxic and cytotoxic compounds (methyl methanesulfonate, ethyl methanesulfonate, etoposide and 4-nitroquinoline-N-oxide) in TK6 human lymphoblast cells. Our data show that measuring ATP and Caspase-3/7 levels provides more rapid and perhaps more biologically relevant measures of cytotoxicity compared with trypan blue dye e...Continue Reading
References
Jan 1, 1992·Mutation Research·M J ArmstrongS M Galloway
Feb 28, 1992·Biochemical and Biophysical Research Communications·K BerghmansJ S Cohen
May 2, 1990·Biochimica Et Biophysica Acta·M NeemanH Degani
Nov 15, 1988·Analytical Biochemistry·L J Kricka
Mar 15, 1993·Journal of Immunological Methods·S P CrouchJ Fletcher
Jun 1, 1993·Mutation Research·M C EliaW W Nichols
Jan 1, 1994·Environmental and Molecular Mutagenesis·M C EliaW W Nichols
Jun 12, 1996·Mutation Research·R D StorerJ G DeLuca
Feb 7, 1997·Toxicology Letters·A Hartmann, G Speit
Jun 1, 1997·Chemical Research in Toxicology·C S NobelA F Slater
Mar 10, 1998·Mutagenesis·L HendersonS Windebank
May 14, 1998·Chemistry & Biology·N A Thornberry
Nov 7, 1998·The Journal of Biological Chemistry·E Dahan-GrobgeldH Degani
Mar 29, 2000·Environmental and Molecular Mutagenesis·R R TiceY F Sasaki
Sep 17, 2002·Cell Death and Differentiation·J R BayascasJ X Comella
Nov 5, 2002·Proceedings of the National Academy of Sciences of the United States of America·Dean A LeMichael A Moskowitz
Sep 4, 2004·The Journal of Biological Chemistry·Danya LiuJacek Hawiger
Sep 10, 2004·Molecular Biology of the Cell·Yan-Guo RenQuinn L Deveraux
Jun 20, 2006·Mutation Research·Paul W HastwellRichard M Walmsley
Nov 14, 2006·Bioorganic & Medicinal Chemistry·Fatih M UckunKatayoon H Emami
Apr 26, 2007·Mutagenesis·Michael D Fellows, Michael R O'Donovan
Feb 20, 2008·Chemical Research in Toxicology·Ruili HuangChristopher P Austin
Feb 20, 2008·Mutagenesis·Andrew R CollinsRudolf Stetina
Oct 22, 2009·Mutagenesis·Jing ShiAnna Szkudlinska
Citations
Oct 19, 2010·Toxicological Sciences : an Official Journal of the Society of Toxicology·Shannon H O'SheaIvan Rusyn
Jan 24, 2012·Toxicological Sciences : an Official Journal of the Society of Toxicology·Eric F LockIvan Rusyn
Oct 19, 2010·Journal of Hazardous Materials·D LiviacR Marcos
Jan 29, 2014·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Natsuko TakakuraLudovic Le Hegarat
Oct 1, 2014·Journal of Toxicology and Environmental Health. Part a·Eşref DemirRicard Marcos
Sep 7, 2012·Mutagenesis·Mike O'Donovan
Jul 19, 2013·Mutagenesis·Aoi KimuraMasamitsu Honma
Apr 12, 2012·Toxicological Sciences : an Official Journal of the Society of Toxicology·Caroline H TophamRichard M Walmsley
Jul 12, 2017·Journal of Applied Toxicology : JAT·Jacky Bhagat
Aug 10, 2018·Environmental and Molecular Mutagenesis·Dejun HuangYongmei Qi
Apr 24, 2014·Environmental and Molecular Mutagenesis·Steven M BryceStephen D Dertinger
Oct 9, 2019·Marine Drugs·S Gill, V M Ruvin Kumara
Jun 29, 2011·Journal of Toxicology and Environmental Health. Part a·Naouale El-YamaniRicard Marcos
Oct 7, 2011·Environmental and Molecular Mutagenesis·Jing ShiSandra Springer