Gene expression profiles of HCC and surrounding non-cancerous tissues in rats fed a CDAA diet for 70 weeks, as well as normal liver tissues, were explored using an oligonucleotide microarray for 3757 genes. A total of 146 genes were identified as differentially expressed; the affected functions including metabolism, apoptosis, cell cycling, RNA splicing, Wnt signaling, reactive oxygen species-induced stress, and fibro/cirrhogenesis. The genes were found to fit into four distinct expression patterns after classification by hierarchical and kmeans clustering procedures. Notably, genes within the same functional category tended to be found within the same cluster, thus gene functions appeared to be related to their expression patterns. For example, genes encoding receptors (Fisher's exact test, P < 0.01) and cytokines (Fisher's exact test, P < 0.05) were both enriched in a cluster characterized by low expression in HCC compared to their surrounding tissues. While some of the receptors in this cluster had cell-proliferative potential, others are known to be growth-suppressive. It was noted, however, that four of the 10 receptor genes encode G-protein-coupled receptors, for which growth-suppressive potential has been reported. The seven growth factors in the same cluster included two fibroblast growth factors. H epatocellular carcinomas are common in Asia and Africa, and the incidence is increasing in Europe and North America. The prognosis for HCC is extremely poor. In human HCC, both genetic and epigenetic alterations have been detected with regard to particular genes such as p53, cyclin D, p16 lnk4 , p21 Waf1/ Cip1 , Rb, β-catenin, mannose-6-phosphate/insulin-like growth factor II receptor (M6P/IGF2R), E-cadherin, cyclo-oxygenase (COX )-2, and telomerase reverse transcriptase (hTERT ).(1-3) In addition to studies on individual genes, microarray technology has allowed the exploration of comprehensive changes in expression during HCC development. To gain insight into the overall picture, suitable animal models are necessary so that samples reflecting various stages of carcinogenesis can be collected. It should be noted, however, that in animal models featuring the use of chemical carcinogens, unavoidable carcinogen-specific molecular alterations may mask generic and essential changes. In addition, the major risk factor in human HCC has been established as being continuous chronic liver injury, which occurs in hepatitis virus infections.(1-3) Thus we have chosen to use an animal model that employs the administration of a CDAA diet, which induces HCC on a background of continuous hepatic injury and cirrhosis. It has been confirmed that this model resembles human carcinogenesis caused by chronic viral hepatitis, hemochromatosis, and Wilson's disease in many respects.(1-4)The CDAA diet is hepatocarcinogenic in male rats of Fischer 344 and Wistar strains.(4) As with other diets deficient in choline and low in methionine (CMD diets), HCC are induced in the absence of chemical carcinogens. In the CDAA model, HCC occur a...