We tested an adenovirus vector that carries a beta-D-galactosidase marker gene for its ability to transduce normal oral mucosa and oral carcinoma cells. Topical application of adenovirus to normal oral mucosa of mice at 1 x 10(10) plaque-forming units (pfu)/mL for 1 minute did not result in transduction of epithelial cells. Similarly, topical application to human oral mucosa ex vivo was not successful. However, systemic administration by intracardiac injection of hamsters did transduce normal oral mucosa effectively. To evaluate transduction of carcinomas, the Tu138 human oral cancer cell line was used. A single application of virus at 1 x 10(8) pfu/mL in vitro resulted in 30% of oral carcinoma cells expressing the marker gene, and 2 x 10(8) pfu/mL transduced 60% of cells. After two applications of virus at 2 x 10(8) pfu/mL with an interval of 18 hours, 100% of oral carcinoma cells expressed the marker gene. Topical application to a raft culture of Tu138 cells for 1 hour produced gene expression that penetrated up to four layers of cells. To emulate the effect of treating a carcinoma, Tu138 cells were implanted subcutaneously in nude mice, allowed to grow to a tumor 1 cm in diameter, and then injected directly with virus. This produced diffuse transduction of around 30% of cells in the tumor, and expression was seen in cells at a significant distance from the injection site. Adenovirus vectors are therefore capable of transferring novel genetic information to both normal and malignant oral mucosa. They may have potential for use in gene therapy in the prevention or treatment of oral squamous carcinomas.