A recent study using c-Ha-ras proto-oncogene transgenic (rasTg) rats demonstrated possible enhancing effects of diacylglycerol (DAG) on 4-nitroquinoline 1-oxide (4NQO) induced carcinogenesis of the tongue. To assess effects in their Sprague-Dawley back strain, a two-stage carcinogenesis study using 4NQO as an initiator was performed. Groups of 30 male rats were initially treated with 4NQO at a dose of 10ppm in the drinking water for the first 10 weeks followed after a 1 week recovery interval by 11% DAG, 5.5% DAG+5.5% triacylglycerol (TAG), 2.75% DAG+8.25% TAG, 1.38 DAG+9.62% TAG, 11% TAG, 11% high linoleic acid TAG (HLTG), 5.5% DAG or 2.75% DAG in the diet for 35 weeks. Further groups of animals were treated with distilled water instead of 4NQO followed by 11% DAG, 11% TAG or 11% HLTG in the same manner. The final survival rates in 4NQO-treated groups were from 50 to 77%. However, incidences and multiplicities of squamous cell papillomas and carcinomas in the oral cavity induced by 4NQO were not affected by any of the dietary treatments. Thus, in contrast to the positive data using rasTg rats, DAG had no potential for enhancing 4NQO-induced tumorigenesis in their back strain.