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Introduction: Hepatocellular carcinoma (HCC) is sixth common cancers worldwide and predominant in Asia and Africa. A number of evidence suggests a possible role of midkine (MDK) and α-fetoprotein (AFP) in the pathogenesis of hepatocellular carcinoma (HCC). Materials and methods: We studied MDK and AFP in patients with HCC or in healthy controls. MDK and AFP was measured by enzyme linked immunosorbent assay and chemiluminescent immunoassay, respectively in 30 patients with primary hepatocellular carcinoma and 30 normal subjects Results: MDK and AFP were found in high levels in the serum of patients initially diagnosed with HCC (18 ± 9.8 pg/ml and 492.01 ± 298.89 pg/ml) compared with healthy subjects (4.29 ± 2.10 pg/ml and 3.13 ± 1.27 pg/ml), respectively. A significant positive correlation was found between mean levels of MDK and AFP in HCC (P < 0.05). Combination of MDK and AFP improved the sensitivity of HCC diagnosis or predicting future HCC development. Conclusion: MDK along with AFP could be considered a promising tumor marker for HCC. In particular, the diagnostic value of the test is significantly increased when combined with AFP.