Podoplanin is a mucin-type glycoprotein widely used as a lymphatic endothelial marker. It is well known that podoplanin is expressed in various neoplasms including oral squamous cell carcinoma (OSCC). Apart from podoplanin expression in cancer cells, recent studies have suggested that podoplanin expression in stromal cancer-associated fibroblasts (CAFs) may be an indicator of poor prognosis in various cancers. In the present study, we performed immunohistochemical analyses of podoplanin and alpha-smooth muscle actin (α-SMA) in OSCC in order to clarify the significance of podoplanin-positive CAFs. Paraffin-embedded tissue specimens of 69 primary and 29 corresponding metastatic lesions in lymph nodes were examined immunohistochemically using antibodies against podoplanin and α-SMA. Podoplanin-positive stromal fibroblasts were detected in 51 (73.9%) of the 69 primary OSCCs and 24 (82.8%) of the 29 lymph nodes metastases. α-SMA immunoreactivity was observed in 39 (56.5%) of the primaries and 24 (82.8%) of the metastases. Further examination showed that 38 (74.5%) of the primary lesions and 23 (95.8%) of the metastases with podoplanin positivity were also positive for α-SMA. In addition, the intensity of α-SMA immunoreactivity increased as that of podoplanin became stronger. Podoplanin-positive CAFs are considered to be myofibroblasts that may contribute to progression of oral cancer.