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

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.

• Authors: Tara C K Krosch, Veena Sangwan, Sulagna Banerjee, Nameeta Mujumdar, Vikas Dudeja, Ashok K Saluja, Selwyn M Vickers
• Journal: American journal of surgery (Am J Surg) Vol. 205 Issue 4 Pg. 387-96 (Apr 2013) ISSN: 1879-1883 [Electronic] United States
• PMID: 23428154 (Publication Type: Evaluation Study, Journal Article, Research Support, N.I.H., Extramural)
• Copyright: Copyright © 2013 Elsevier Inc. All rights reserved.

Chemical References

• Antineoplastic Agents, Alkylating
• Biomarkers, Tumor
• Diterpenes
• Epoxy Compounds
• Heat-Shock Proteins
• MAP1LC3A protein, human
• Microtubule-Associated Proteins
• NF-kappa B
• Phenanthrenes
• triptolide
• Caspase 3
• Caspase 7
• Calcium

Topics

• Antineoplastic Agents, Alkylating (pharmacology, therapeutic use)
• Apoptosis (drug effects)
• Autophagy (drug effects)
• Biomarkers, Tumor (antagonists & inhibitors, metabolism)
• Blotting, Western
• Calcium (metabolism)
• Caspase 3 (metabolism)
• Caspase 7 (metabolism)
• Cell Line, Tumor
• Cell Survival (drug effects)
• Diterpenes (pharmacology, therapeutic use)
• Dose-Response Relationship, Drug
• Epoxy Compounds (pharmacology, therapeutic use)
• Fluorescent Antibody Technique
• Heat-Shock Proteins (metabolism)
• Humans
• Microtubule-Associated Proteins (metabolism)
• NF-kappa B (antagonists & inhibitors)
• Neuroblastoma (drug therapy, metabolism)
• Phenanthrenes (pharmacology, therapeutic use)
• Real-Time Polymerase Chain Reaction