Abstract: Objective To investigate the effect of lithium chloride (LiCl) on cell cycle of HK-2 cells and explore the possible pathways involved. Methods HK-2 cells were treated with LiCl at different concentrations (5, 12.5, 20, and 25 mmol/L) for 12, 24, 48, or 72 h, and the changes in cell cycle and viability were detected using flow cytometry and CCK-8 assay, respectively. Western blotting was used to analyze the changes in the expressions of cyclin B1 and CDK1 (the two G2 phase-related proteins) and those of AKT/GSK-3β signaling pathway-related proteins in the treated cells. Results LiCl treatment time- and concentration-dependently increased HK-2 cell percentage in G2 phase and decreased the cell vitality. The expressions of cyclin B1, CDK1, p-GSK-3β, and β-catenin increased and the expression of p-AKT decreased significantly in the cells as LiCl treatment time and concentration increased. Conclusion LiCl may cause HK-2 cell cycle arrest in G2 phase through activation of the AKT/GSK-3β signaling pathway.