Homeoprotein Six1 belongs to a subfamily of the Six family of homeodomain-containing transcription factors that shares a lysine within the DNA-binding helix of the homeodomain. 6, 7 Six1, located at 14q23 of the chromosome, is involved in early developments of diverse organs such as the brain, ear, eye, muscle and kidney. 7–11 Alteration of Six1 expression takes place in human breast cancer, Wilms’ cancer, cervical cancer, ovarian cancer and rhabdomyosarcoma, indicating its possible contributions in the tumorigenicity of different cancers. 12–16 Six1 can transform the adult mammary gland to hyperplasia and highly aggressive tumor as well as a stem cell phenotype suggesting an oncogenic role of Six1. 17 Six1 is also regarded as an important metastatic regulator in cancers. For example, overexpression of Six1 occurs in a large percentage of primary cancers and strongly correlates with breast lesions. 14 Six1 elevated in breast cancer promotes progression of breast cancer. 18 Overexpression of Six1 promotes metastasis of breast cancer through amplifying TGF-b signaling and inducing an epithelial-mesenchymal transition (EMT). 19 Six1 also plays a substantial role in regulating the metastatic ability of rhabdomyosarcoma. 12 The impact of Six1 on promoting tumorigenesis and metastasis of multiple cancers therefore impelled us to study the property of Six1 in HCC. We previously found that Six1 protein is specifically overexpressed in tumor tissues rather than nontumor tissues