Background Hepatocellular carcinoma (HCC) is a major unresolved medical issue, which is considered as the seventh most common cancer globally and the fourth leading cause of cancer deaths, with approximately 700,000 deaths per year. Aim of the Work The aim of this study; to evaluate the role of serum vimentin as a diagnostic biomarker in patients with HCC. Patients and Methods This case control study enrolled 60 patients; divided into two groups: Group (I): included 20 patients with liver cirrhosis without HCC as a control group and Group (II): 20 patients divided into 2 subgroups: (IIa): includes 20 patients with HCC and serum AFP is high and (IIb): includes 20 patients with HCC and serum AFP is normal. Results The present study found statistical significant decrease in Hb level in group IIb compared to group I, there was high statistical significant decrease in platelet count in patients of group IIa and IIb compared to control group. a statistical significant elevation of total bilirubin and INR with decrease of albumin in group II compared to control group.There was high statistical significant elevation in AFP in group IIa and IIb compared to control group and in group IIa compared to group IIb, a high statistical significant elevation in Vimentin level in group IIa and IIb compared to control group while there was no statistical significant difference between group IIa and group IIb. Conclusion Vimentin can be considered as an important independent and discriminative serological biomarker for detecting early stage HCC in patients with normal serum AFP.