Although the diagnostic and therapeutic modalities of esophageal squamous cell carcinoma (ESCC) have been improved considerably, the five-year survival rate is still not satisfied. To detect the numberial aberrations of the chromosomes in ESCC, fluorescence in situ hybridization (FISH) was performed on interphase nuclei prepared from 220 esophageal carcinoma tissues with specific centromeric probes for chromosomes 3, 8, 10, 20 and Y. The main aberrations of the euchromosomes was chromosome gain, including trisome, tetrasome, and polysome. The gain rates of the four euchromosome were 84.9%, 77.5%, 63.7% and 83.2%, and the frequencies of polysome for each euchromosome were 24.6%, 34.9%, 23.4% and 31.7%, respectively. Loss of chromosome Y was observed in 61.2% of male patients. The combination of the four chromosome probes 3, 8, 10 and 20 detected 74.5% of ESCC and the combination of 3, 8, 20 and Y detected 85.0%. These results indicated that both sets of the four centromeric probe combinations provide candidate biomarkers for the diagnosis of esophageal squamous cell carcinoma.