Triptolide-mediated cell death in neuroblastoma occurs by both apoptosis and autophagy pathways and results in inhibition of nuclear factor–kappa B activity

• Published in: The American journal of surgery Vol. 205; no. 4; pp. 387 - 396
• Main Authors: Krosch, Tara C.K., M.D, Sangwan, Veena, Ph.D, Banerjee, Sulagna, Ph.D, Mujumdar, Nameeta, Ph.D, Dudeja, Vikas, M.D, Saluja, Ashok K., Ph.D, Vickers, Selwyn M., M.D
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.04.2013; Elsevier Limited
• Subjects: Antineoplastic Agents, Alkylating - pharmacology; Antineoplastic Agents, Alkylating - therapeutic use; Apoptosis; Apoptosis - drug effects; Autophagy; Autophagy - drug effects; Biomarkers, Tumor - antagonists & inhibitors; Biomarkers, Tumor - metabolism; Blotting, Western; Calcium - metabolism; Caspase 3 - metabolism; Caspase 7 - metabolism; Cell culture; Cell Line, Tumor; Cell Survival - drug effects; Diterpenes - pharmacology; Diterpenes - therapeutic use; Dose-Response Relationship, Drug; Epoxy Compounds - pharmacology; Epoxy Compounds - therapeutic use; Fluorescent Antibody Technique; Heat shock protein; Heat shock proteins; Heat-Shock Proteins - metabolism; Humans; Microtubule-Associated Proteins - metabolism; Neuroblastoma; Neuroblastoma - drug therapy; Neuroblastoma - metabolism; NF-kappa B - antagonists & inhibitors; NF-κB; Pancreatic cancer; Pediatrics; Phenanthrenes - pharmacology; Phenanthrenes - therapeutic use; Real-Time Polymerase Chain Reaction; Rodents; Studies; Surgery; Triptolide; Tumors; NF-κB; Neuroblastoma; Autophagy; Triptolide; Heat shock protein; Apoptosis
• ISSN: 0002-9610; 1879-1883
• DOI: 10.1016/j.amjsurg.2013.01.008

Abstract Background Neuroblastoma is an aggressive pediatric malignancy with significant chemotherapeutic resistance. We assessed triptolide as a potential therapy. Methods SH-SY5Y and IMR-32 neuroblastoma cell lines were treated with triptolide. Viability, intracellular calcium, caspase activation, protein, and mRNA levels were measured. Autophagy was evaluated with confocal microscopy. Nuclear factor–kappa B (NF-κB) activation was measured using a dual luciferase assay. Results Triptolide treatment resulted in death in both cell lines within 72 hours, with sustained increases in intracellular calcium. IMR-32 cells underwent cell death by apoptosis. Conversely, light chain 3II (LC3II) protein levels were elevated in SH-SY5Y cells, which is consistent with autophagy. Confocal microscopy confirmed increased LC3 puncta in SH-SY5Y cells compared with control cells. Heat shock pathway protein and mRNA levels decreased with treatment. NF-κB assays demonstrated inhibition of tumor necrosis factor (TNF)-α–induced activity with triptolide. Conclusions Triptolide treatment induces cell death in neuroblastoma by different mechanisms with multiple pathways targeted. Triptolide may serve a potential chemotherapeutic role in advanced cases of neuroblastoma.