Glycan-binding protein play important biological roles in biological processes. We use carbohydrate microarray to study the alteration of GBP in hepatocellular carcinoma cell line HepG2 and L02. Carbohydrate histochemistry was used to further validate the GBP and assess the distribution. As a result, 8 carbohydrate probes (e. g. SL, LNT, and GalNAc) showed increased signal while 5 carbohydrate probes (e. g. Man, Man-9-Glycan, and Xyl) showed decreased signal in HepG2 compared with L02 cell line. Meanwhile, GalNAc staining showed moderate binding to the cytoplasma membrane, central cytoplasm, and perinuclear cytoplasm in the L02, and the binding intensified in the same regions of the HepG2. NeuAc staining showed moderate binding to the cytoplasma membrane, and perinuclear cytoplasm in the HepG2, and the binding intensified in the same regions of the L02. In conclusion, the precision alteration of GBP related to HepG2 may provide useful information to find new molecular mechanism of hepatocellular carcinoma and antitumor therapeutic strategies.