Abstract：Objective To study the role of canagliflozin in inhibiting galectin-3-mediated proliferation and migration of intrahepatic cholangiocarcinoma cells. Methods TIMER2.0, GEPIA2 and R2 databases were used to analyze the expression of Gal-3 in cholangiocarcinoma and whether it had any effect on the overall survival rate of cholangiocarcinoma patients. The expression of Gal-3 in intrahepatic cholangiocarcinoma tissues and paracarcinoma tissues was detected by immunohistochemistry. Molecular docking was used to analyze the binding of canagliflozin to Gal-3. Cell experiments were performed with the intrahepatic cholangiocarcinoma cell line HUCCT1, and cells were treated with different concentrations of canagliflozin for 24h and 48h, and the effect of canagliflozin on cell survival rate was detected by CCK8. After treating cells with 16μM canagliflozin for 24h, the effect of canagliflozin on the proliferation and migration of cells was analyzed by using EdU Fluorescent Labeling and the Wound-Healing Assay. Results Bioinformatics Analysis results showed that Gal-3 was significantly highly expressed in cholangiocarcinoma, and the immunohistochemical results using intrahepatic cholangiocarcinoma tissues and paracarcinoma tissues also showed that the expression of Gal-3 was significantly increased in intrahepatic cholangiocarcinoma, and that the high expression of Gal-3 was significantly correlated with the overall survival rate of the cholangiocarcinoma patients. The results of the molecular docking study revealed that canagliflozin is capable of recognizing Gal-3 at a novel site instead of the conventional carbohydrate binding site. As the concentration of canagliflozin treatment increased and the treatment time was prolonged, the cell survival rate decreased. After treating HUCCT1 cells with 16μM canagliflozin for 24 h, the results of EdU Fluorescent Labeling showed that the proliferative ability of the treated group was significantly lower than that of the control group. The results of the Wound-Healing Assay showed that the cell migration rate of the treated group was significantly lower than that of the control group. Conclusion Canagliflozin inhibits the proliferation and migration of intrahepatic cholangiocarcinoma cells by binding to Gal-3.
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