Objective: To investigate the relationship between p16INK4a and primary hepatocellular carcinoma (HCC),especially hepatitis B-related HCC. Methods: p16INK4a and its protein in HCC were analyzed with PCR-SSCP and the immunohistochemistry methods respectively. Results: The positive incidence of p16INK4 protein expressing in HCC was lower than that of normal liver tissue (P＜0.05), and the absence of p16INK4 protein was associated with HCC metastasis (P＜0.05). The low frequency of mutation of p16INK4 exonl and exon2 upstream fragment was found in HCC. Conclusion: Absence of p16INK4 protein in HCC was not associated with HBV-infection.