C4.4A is a glycolipid-anchored membrane protein expressed in several human malignancies. We recently found that C4.4A expression was associated with poor prognosis of esophageal squamous carcinoma cells (ESCCs), but the underlying mechanism is unknown. To uncover this, we performed PCR array analysis using the HCT116 cell line, a positive control for C4.4A expression and we found that Tenascin-C (TNC) among the many adhesion molecules and extracellular matrix proteins was the best candidate for C4.4A molecule induction. Based on in vitro studies using the TE8 esophageal cancer cells, we examined by immunohistochemistry TNC expression in 111 ESCCs. We found that the TNC-positive group (24.3%) had significantly poorer prognosis than the TNC-negative group in 5-year overall survival. We also found there was a significant correlation between TNC and C4.4A in ESCC tissues (P=0.007). Finally, we found that only the double-positive group for C4.4A and TNC had a significantly worse prognosis (P=0.005). Our data suggest that TNC expression in ESCC may in part explain why C4.4A is associated with a poor prognosis of ESCC since TNC can promote invasion and metastasis.