We investigated the interaction between tumor cells and stromal fibroblasts in tumor invasion of oral squamous cell carcinoma. Gelatin zymography showed that high levels of matrix metalloproteinase (MMP)-9 were present in the tissue of squamous cell carcinoma. When tumor cells and fibroblasts were isolated from the tissue and cultured separately, significant levels of MMP-9 were lost in the culture media of tumor cells as well as fibroblasts. When tumor cells and fibroblasts were cocultured in the presence of tumor necrosis factor alpha, high levels of MMP-9 were recovered in the culture media. The levels of MMP-9, which were secreted from tumor cells, but not fibroblasts, correlated with the number of cocultured fibroblasts. Cocultured fibroblasts, moreover, enhanced the induction of an active form of MMP-9, cell motility, and the activation of a transcription factor NF-kappaB in tumor cells. Stromal fibroblasts may induce NF-kappaB activation and promote the invasion of oral squamous cell carcinoma.