Gomisin N enhances TRAIL-induced apoptosis via reactive oxygen species-mediated up-regulation of death receptors 4 and 5

Pharmacological studies have revealed that Lignans isolated from Schisandra chinensis, including gomisin N, show Anticancer, anti-hepatotoxic, anti-oxidative and anti-inflammatory activities. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is an important member of the tumor necrosis factor superfamily with great potential in Cancer therapy. The present study investigated whether pretreatment with gomisin N significantly enhanced TRAIL-induced cleavage of Caspase-3, Caspase-8 and PARP-1, which are key markers of Apoptosis. Pretreatment with z-VAD-FMK, a pan-caspase inhibitor, was able to inhibit Apoptosis enhanced by the combination of gomisin N and TRAIL. These results suggested that gomisin N could promote TRAIL-induced Apoptosis through the Caspase cascade. In search of the molecular mechanisms, we elucidated that such enhancement was achieved through transcriptional up-regulation of TRAIL receptors, Death Receptor 4 (DR4) and DR5. Neutralization of DR4 and DR5 could significantly reduce Apoptosis induced by gomisin N and TRAIL. We also revealed that gomisin N increased the generation of Reactive Oxygen Species (ROS). N-acetyl cysteine (NAC), an antioxidant, could inhibit ROS production and up-regulation of DR4 and DR5. Overall, our results indicated that gomisin N was able to potentiate TRAIL-induced Apoptosis through ROS-mediated up-regulation of DR4 and DR5.