Background: In Egypt, hepatocellular carcinoma (HCC) is a disease with increased prevalence rate and a dismal prognosis and survival rate. For an early diagnosis of HCC, biomarkers are crucial. The glycoprotein osteopontin (OPN), which is released by T cells, osteoblasts, and macrophages, is also strongly expressed in a wide range of tumors, including stomach and colon. Aim: Our aim was to evaluate the serum level of OPN as a potential marker for HCC patients with HCV, in comparison with other tumor markers. Materials and Methods: This study included 140 patients (aged 27-79 years) divided into four groups. Group I involved 20 healthy controls, Group II involved 40 patients with HCV, Group III involved 40 patients with HCC. Group IV involved 40 patients with HCV and HCC. Routine clinical tests such as ALT, AST, ALP, ALBUMIN and BILIRUBIN were assayed for all patients. In addition, serum level of OPN was identified using Western blotting and its levels were quantified using sandwich ELISA. The absolute values of the investigated markers were statistically analyzed using SPSS program. Results: The mean OPN level was 11.89±7.5, 47.34 ±2. 71, 201 ±5.85, and 251.52± 19.63 pg/ml for G1, G2, G3 and G4, respectively. Of interest, Group IV (patients with HCC and HCV) showed much higher serum OPN levels than the normal group (p=0.001). Using Western blotting assay, OPN was identified at 68-kDa molecular mass in the sera from all patients. Conclusion: OPN has a greater sensitivity and specificity than CA.CA19.9 and AFP. It useful for early detection of HCC.