Objective To investigate the expressions of nucleostemin(NS), epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) mRNA and their relationship in human esophageal squamous cell carcinoma(ESC) tissue. Methods The expressions of NS, EGF and EGFR mRNA in 62 cases of esophageal squamous cell carcinoma, 31 cases of dysplasia and 62 cases of normal esophageal tissue were determined by EM>in situ/EM> hybridization. The relationship of NS, EGF and EGFR mRNA with the tumor grading, the infiltration and lymphatic metastasis of ESC was analyzed. Results In the normal esophageal tissue, dysplasia esophageal tissue and ESC tissue, the positive expression rates of NS mRNA were 21.0% (13/62), 25.8% (8/31) and 69.4% (43/62) respectively; the positive expression rates of EGF mRNA were 40.3% (25/62), 48.4% (15/31) and 77.4% (48/62); the positive expression rates of EGFR mRNA were 30.6% (19/62), 45.2% (14/31) and 75.8% (47/62). The expressions of NS, EGF and EGFR mRNA showed higher positive correlation with the tumor grading, the infiltration and lymphatic metastasis of ESC (all P<0.05). The expression of NS mRNA was positively correlated with that of EGF and EGFR mRNA (EM>r/EM>=0.394 and EM>r/EM>=0.604, EM>P/EM><0.05)in ESC. Conclusion The NS, EGF and EGFR mRNA may play an important role in the genesis and progression of ESC.