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 Sarcophyton crassocaule 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 mu M). 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 mu M) 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 mu M 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%similar to 5.4%/22.7% for HA22T cells, in the HePG2 cells were 0.6%/0.2%similar to 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 p-4EBP1 increased with an increasing concentration of 13-acetoxysarcocrasside (0, 1, 2, and 4 mu M), whereas that of Bcl-2, Bcl-xL, Mcl-1, p-Bad, p-PI3K, p-AKT, p-mTOR, p-70S6K, p-S6, p-eIF4E, and p-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/p70S6K pathway. The potential of 13-acetoxysarcocrassolide as a chemotherapeutic agent should be further assessed for use in human hepatocellular carcinoma treatment.

Automated summary

The study found that 13-acetoxysarcocrassolide, extracted from Taiwanese soft coral, could induce apoptosis in hepatocellular carcinoma cells through mitochondrial dysfunction and inactivation of the PI3K/AKT/mTOR/p70S6K pathway. This research is of great significance for evaluating the potential of 13-acetoxysarcocrassolide as a therapeutic agent for hepatocellular carcinoma.