Epidemiologic and clinical evidence suggests a possible relationship between the human papillomavirus types found in oral squamous cell carcinoma and cervicovaginal cancer. To try to address this question, we analyzed by in situ hybridization 50 paraffin-embedded biopsies of oral squamous cell carcinomas and their corresponding Papanicolaou smears using nonradioactive DNA probes for human papillomavirus type 6/11 and 16/18. This analysis revealed that 70% (35 of 50) of oral squamous cell carcinomas and 56% (28 of 50) of Papanicolaou smears were positive for one or both of these human papillomavirus DNA probes. In 23 of the 28 patients who were positive for human papillomavirus sequences in the cervicovaginal tract, we found the same human papillomavirus type in the oral cavity. Five of the positive in situ hybridization Papanicolaou smears were associated with negative in situ hybridization oral squamous cell carcinomas. Our data suggest that after a primary infection by human papillomavirus, other mucosal sites may eventually become infected. Local environmental factors may contribute to the pathogenesis of lesions such as oral squamous cell carcinoma.