The majority of studies report that liver kinase B1 (LKB1) acts as a tumor suppressor by inhibiting cell proliferation and metastasis. The present study investigated the expression pattern of LKB1 in 2 cohorts of paired hepatocellular carcinoma (HCC) and analogous non-cancerous tissues (ANT). The results indicated that LKB1 was upregulated in HCC vs. ANT tissues, and that high expression of LKB1 was associated with a higher number of tumor foci, larger tumor size, poorer tumor differentiation, Edmondson-Steiner grade, Barcelona Clinic Liver Cancer grade and tumor-node-metastasis stage. Furthermore, high LKB1 expression was associated with poor overall survival (OS), shorter disease-free survival and early recurrence. Univariate and multivariate analyses demonstrated that high LKB1 expression may serve as an independent prognostic marker for OS, but not for recurrence. In addition, knockdown of LKB1 expression in HCC cell lines inhibited cell proliferation and subcutaneous tumor growth by promoting cell apoptosis. Therefore, the findings of the present study suggest a protooncogenic role of LKB1 in HCC.