Subsequent quantitative RT‐PCR confirmed that compared with parental Huh‐7 cells, Huh‐7‐trans and Huh‐7‐DN subpopulations lost mature hepatocyte‐specific biomarkers (AFP, CEBPA, CYP2C9, UGT1A1, GSTA1/2) and significantly increased expression of cancer progression‐associated genes (FGF1/5, PDGFB, PDGFRB, TGFB1, VEGFC, WNT5B), indicating that these two subpopulations were poorly differentiated. This evidence concerns the gene AFP and cancer.