We examined AIS status in digestive tract cancers and found that all eight esophageal cancer cell lines (100%) showed AIS/TA-AIS gene overexpression, whereas 1 of 12 (8%) gastric cancer and 0 of 14 (0%) colon cancer cell lines showed AIS/TA-AIS gene expression. We then confirmed that the AIS gene, not the TA-AIS gene, was dominantly expressed in esophageal cancers by reverse transcription-PCR. AIS protein was also expressed in AIS gene-positive cell lines. Subsequently, we tested AIS gene expression in paired esophageal normal tissues and cancers. Twenty-five of 39 (64%) primary esophageal cancers demonstrated an obviously higher expression of AIS gene compared to paired normal tissues. Moreover, high AIS gene expression was significantly associated with lymph node metastases in esophageal cancer (P = 0.0271). These results suggested that AIS may be useful as a marker for advanced esophageal cancer.