Abstract: AIM The present study aimed to determine the role of sphingosine kinase-2 (SphK2) in angiotensin-II (AngII)-induced cardiomyocyte hypertrophy. METHODS Cardiomyocytes were isolated from Sprague Dawley rats and were cultured in vitro. Then, cardiomyocytes were exposed to AngII (10 μmol/L) for 24 hours. The expression of SphK2 was determined by Western blot and nuclear S1P abundance was analyzed by enzyme-linked immunosorbant assay (ELISA). After vehicle or ABC294640 (1 μmol/L) co-treatment, AngII-induced cardiomyocyte hypertrophy was assayed by crystal violet staining. The mRNA expression levels of hypertrophy marker genes, including artial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and β-myosin heavy chain (β-MHC), were determined by real-time polymerase chain reaction (RT-PCR). RESULTS The AngII treatment promoted cardiomyocyte hypertrophy and upregulated expression of hypertrophy marker genes. ABC294640 treatment significantly reduced nuclear S1P abundance in the cardiomyocyte. Notably, compared with the vehicle group, ABC294640 aggravated AngII-induced cardiomyocyte hypertrophy, as evidenced by increased cardiomyocyte size and expression of hypertrophy marker genes. CONCLUSION Inhibition of SphK2 exacerbates AngII-induced cardiomyocyte hypertrophy. SphK2 may be a potential therapeutic target for the prevention and treatment of pathological cardiac hypertrophy.