Hepatocellular carcinoma (HCC) is one of the common malignant tumors in China. Chronic hepatitis B virus (HBV) infection has been considered as a risk factor for HCC. Hepatitis B virus X protein (HBx) is a multifunctional regulator that regulates signal transduction and epigenetic gene expressions, and also is involved in cell proliferation and apoptotic pathways. HBx can promote the self-renewal ability, differentiation, and resistance to chemotherapy and other biological behaviors of the liver cancer stem cells (LCSCs), and thereby promote tumor metastasis, infiltration and recurrence. The authors address HBx regulating LCSCs to maintain the biological behaviors of HCC.