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The esophageal squamous cell carcinoma (ESCC) has high incidence in Shaanxi Province of China. More and more researches indicated that human papillomavirus type 16 (HPV16) might play an important role in carcinogenesis of ESCC but the relationship between HPV16 and CD44v6, nm23H1 has not been elucidated. HPV16 was detected by amplifying HPV16 E6 gene through polymerase chain reaction (PCR) method and the expression of CD44v6, nm23H1 in 40 ESCCs and fifteen normal esophageal mucosa (NEM) from Shaanxi Province was examined by Streptavidin-Peroxidase (SP) method using monoclonal antibody specific to CD44v6 and nm23H1. The positive rates of HPV16 E6 gene, CD44v6 and nm23H1 were 60% (24/40), 65% (26/40) and 45% (18/40) respectively in ESCCs and 26.67% (4/15), 33.33% (5/15) and 86.67% (13/15) respectively in NEMs. There exited statistical difference for HPV16, CD44v6 and nm23H1 between NEMs and ESCCs respectively (p<0.05). The relationship between HPV16 and the expression of CD44v6 in ESCCs was statistical significance (P?=?0.021), but no significant correlation was found between HPV16 and the expression of nm23H1 (P?=?0.436) in ESCCs. The infection rate of HPV16 had no statistical difference in all pathological features we observed, but the expression rates of CD44v6 and nm23H1 had statistical correlation with invasion (p?=?0.001, 0.013) and lymph nodes metastasis (p?=?0.014, 0.002) respectively. In different histology grade of ESSCs, the relationship between HPV16 and CD44v6 was statistical significance in grade I (p?=?0.044) but was not in grade II (p?=?0.165) and grade III (p?=?0.658), however as to the expression of nm23H1 there exited no statistical significance in all histology grades of ESCC (p?>?0.05). The expression rates of CD44v6 and nm23H1 were statistically different between grade I and II (p?=?0.004, 0.016) respectively and between grade I and grade III (p?=?0.014, 0.020), but not statistically different between grade II and III (p?=?0.792, 0.943) respectively. Our data firstly suggested that there existed the statistical relationship between the infection of HPV16 and the expression of CD44v6 in ESCCs and that HPV16 may be involved in invasion and metastasis of ESCC. [ABSTRACT FROM AUTHOR]