Therapeutic Discovery DLK1 as a Potential Target against Cancer Stem/Progenitor Cells of Hepatocellular Carcinoma

Delta-like 1 homolog (DLK1;Drosophila) is a hepatic stem/progenitor cell marker in fetal livers that plays a vital role in oncogenesis of hepatocellular carcinoma (HCC). The aim of this study is to investigate whether DLK1 could serve as a potential therapeutic target against cancer stem/progenitor cells of HCC. DLK1þ and DLK1 cells were sorted by fluorescence-activated cell sorting and magnetic-activated cell sorting, respectively, and then were evaluated by flow cytometry. The biological behaviors of these isolated cells and those with DLK1 knockdown were assessed by growth curve, colony formation assay, spheroid colony formation, chemoresistance, and in vivo tumorigenicity. Adenovirus-mediated RNA interferencewas used to knockdown the endogenous DLK1. We found that DLK1þ population was less than 10% in almost all 17 HCC cell lines examined. DLK1þ HCC cells showed stronger ability of chemoresistance, colony formation, spheroid colony formation, and in vivo tumorigenicity compared with DLK1 cells. The DLK1þ HCC cells could generate the progenywithoutDLK1 expression. Furthermore,DLK1 knockdown could suppress the ability of proliferation, colony formation, spheroid colony formation, and in vivo tumorigenicity of Hep3B and Huh-7 HCC cells. Our data suggested that DLK1þ HCC cells have characteristics similar to those of cancer stem/progenitor cells. RNA interference againstDLK1 can suppress the malignant behaviors of HCC cells, possibly through directly disrupting cancer stem/progenitor cells, which suggested that DLK1 could be a potential therapeutic target against the HCC stem/progenitor cells. Mol Cancer Ther; 11(3); 629–38. 2012 AACR.