Sponge Dysidea Avara induction of ROS mediated apoptosis in human glioblastoma cells via mitochondrial targeting

Sponge Dysidea Avara induction of ROS mediated apoptosis in human glioblastoma cells via mitochondrial targeting
Mohammad Reza Neshat,^1,* Davar Mohammadpour,² Jalal Pourahmad,³
1. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences
2. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences
3. Department of Pharmacology and Toxicology, Faculty of Pharmacy, Shahid Beheshti University of Medical Sciences
Introduction: Glioblastoma is the most lethal brain tumor with poor prognosis which possesses a high resistance against anticancer drugs. Marine organisms are an abundant source of bioactive molecules. Different studies on marinesponges indicated that sponge Dysidea avara, have cytotoxic effects on cancer cell lines; therefore it is suggested that marine drugs can potentially be used as a beneficialmedicine in cancer therapy. Sponge Dysidea Avara’s anti-cancer effects and associated mechanisms were assessed in brain tissues, focusing on parameters of inflammatory change and apoptosis.
Methods: Mitochondria were isolated from the glioblastoma cells by mechanical lysis and multiple centrifugations. The activity of mitochondrial complex II was assayed via the measurement of MTT reduction. The mitochondrial ROS measurement was performed using the fluorescent probe DCFH-DA. The Rhodamine 123 (Rh 123) redistribution technique was used for MMP measurement. Mitochondrial swelling was measured spectrophotometrically in duration 1 hour. Caspase-3 activity was evaluated using the Sigma caspase-3 assay kit. Data were analyzed using the Graph pad prism software, version 7.
Results: Our results demonstrated that Dysidea Avara induced a rise in mitochondrial reactive species (ROS) formation and mitochondrial membrane potential (MMP) collapse before mitochondrial swelling ensued in isolated brain mitochondria. In addition collapse of MMP and mitochondrial swelling produced release of cytochrome c via outer membrane rupture or mitochondrial permeability transition (MPT) pore opening. Furthermore, caspase-3 activity was significantly increased in cells isolated from the brain when incubated with Dysidea Avara.
Conclusion: The present study concluded that Dysidea Avara could be a potential anticancer agent. However, additional studies are needed to clarify involved mechanisms.
Keywords: Apoptosis, Dysidea Avara, Glioblastoma, Mitochondria