13-Acetoxysarcocrassolide induces apoptosis in human hepatocellular carcinoma cells through mitochondrial dysfunction and suppression of the PI3K/AKT/mTOR/p70S6K signalling pathway.

CONTEXT

13-Acetoxysarcocrasside, isolated from the Taiwanese soft coral Moser (Alcyoniidae), has biological activity and induces apoptosis in hepatocellular carcinoma cells.

OBJECTIVE

To elucidate the mechanisms underlying apoptosis induced by 13-acetoxysarcocrasside in HA22T and HepG2 hepatocellular carcinoma cells.

MATERIAL AND METHODS

MTT and morphology assays were employed to assess the anti-proliferative effects of 13-acetoxysarcocrasside (1-5 μM). TUNEL/DAPI staining and annexin V-fluorescein isothiocyanate/propidium iodide staining were used to detect apoptosis. Cells were treated with13-acetoxysarcocrassolide (0, 1, 2, and 4 μM) for 24 h, and the mechanism of cells apoptotic was detected by western blotting. Cells treated with DMSO were the control.

RESULTS

Survival of the cells decreased with the addition of 13-acetoxysarcocrassolide, and at 4 μM cell survival was inhibited by approximately 40%. After treatment of cells with 13-acetoxysarcocrassolide, the incidence of early/late apoptosis to be 0.3%/0.5%∼5.4%/22.7% for HA22T cells, in the HePG2 cells were 0.6%/0.2%∼14.4%/23.7%. Western blotting analysis showed that the expression of Bax, Bad, cleaved caspase 3, cleaved caspase 9, cleaved-PARP-1, cytochrome c, and -4EBP1 increased with an increasing concentration of 13-acetoxysarcocrasside (0, 1, 2, and 4 μM), whereas that of Bcl-2, Bcl-xL, Mcl-1, -Bad, -PI3K, -AKT, -mTOR, -70S6K, -S6, -eIF4E, and -eIF4B decreased.

DISCUSSION AND CONCLUSIONS

Apoptosis induced by 13-acetoxysarcocrassolide in HA22T and HepG2 cells is mediated by mitochondrial dysfunction and inactivation of the PI3K/AKT/mTOR/70S6K pathway. The potential of 13-acetoxysarcocrassolide as a chemotherapeutic agent should be further assessed for use in human hepatocellular carcinoma treatment.