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Molecular profiling of non-genotoxic hepatocarcinogenesis using differential display reverse transcription-polymerase
J C Rockett1, D J Esdaile, G G Gibson
1Molecular Toxicology Group, School of Biological Sciences, University of Surrey, Guildford, UK.
Abstract:
The technique of differential display reverse transcription-polymerase chain reaction (ddRT-PCR) has been used to produce unique profiles of up-regulated and down-regulated gene expression in the liver of male Wistar rats following short term exposure to the non-genotoxic hepatocarcinogens, phenobarbital and WY-14,643. Animals were treated for 3 days, whereupon their livers were extracted and snap frozen. mRNA was prepared from the livers and used for ddRT-PCR. Individual bands from the differential displays were extracted and cloned. False positives were eliminated by dotblot screening and true positives then sequenced and identified.