We report on the cloning and characterization of a novel gene EC97 which is associated with human esophageal squamous cell carcinoma (ESCC). A fragment of expressed sequence tag (EST) (aa700351) was overexpressed in ESCC tissues compared with the normal tissues in cDNA microarray data. Based on the sequence of aa700351, the full-length cDNA was acquired by polymerase chain reaction (PCR) amplification and named EC97. EC97 was 3353 bp long and its encoding protein contained 813 amino acid residues. Northern blot and reverse transcription polymerase chain reaction (RT-PCR) analysis revealed that EC97 had a transcript of 3.4 kb in most human normal tissues we checked and its expression level was increased in 60% (12/20) of tested ESCC tissues versus the normal counterparts. EC97 was mapped to human chromosome 16p12-16p13.1 using radiation hybridization (RH) and predicted to include 25 exons and disseminated over 100 kb of genomic DNA.