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Updated: Jul 2, 2025
Author Spotlight: Tracing the Ferroptotic Signatures and Cell Death Dynamics in Medulloblastoma for Advanced Therapeutics
Published on: March 15, 2024
IP3R-Mediated Calcium Release Promotes Ferroptotic Death in SH-SY5Y Neuroblastoma Cells
Joaquín Campos1, Silvia Gleitze2, Cecilia Hidalgo2,3,4
1Chica and Heinz Schaller Foundation, Institute for Anatomy and Cell Biology, University of Heidelberg, 69120 Heidelberg, Germany.
Ferroptosis, an iron-dependent cell death, involves calcium release via IP3R channels in SH-SY5Y cells. GPx4 inhibition triggers this, increasing calcium, ROS, and lipid peroxidation, ultimately causing cell death.
Area of Science:
- Cell Biology
- Neuroscience
- Biochemistry
Background:
- Ferroptosis is an iron-dependent cell death pathway characterized by glutathione depletion and lipid peroxidation.
- Inhibition of glutathione peroxidase 4 (GPx4) is a known inducer of ferroptosis.
- Calcium release from the endoplasmic reticulum (ER) via ryanodine receptors (RyRs) has been implicated in ferroptosis in neurons.
Purpose of the Study:
- To investigate the role of inositol 1,4,5-trisphosphate receptor (IP3R) channels in ferroptosis-induced cell death.
- To determine if IP3R-mediated calcium release contributes to ferroptosis in SH-SY5Y neuroblastoma cells, which lack RyR channels.
Main Methods:
- Treatment of SH-SY5Y cells with RSL3, a GPx4 inhibitor.
- Inhibition of IP3R channels using Xestospongin B or carbachol.
- Knockdown of IP3R1 expression.
- Intracellular calcium chelation with BAPTA-AM.
- Assessment of reactive oxygen species (ROS) generation, cytoplasmic and mitochondrial calcium levels, lipid peroxidation, and cell morphology.
Main Results:
- RSL3 treatment induced ferroptosis, evidenced by increased ROS, calcium levels, and lipid peroxidation.
- Inhibition of IP3R channels (using Xestospongin B, carbachol, or IP3R1 knockdown) significantly reduced RSL3-induced calcium signals and prevented cell death and morphological changes.
- RSL3-induced calcium signals were dependent on intracellular calcium release, not extracellular calcium.
Conclusions:
- GPx4 inhibition activates IP3R-mediated calcium release from the ER in SH-SY5Y cells.
- This calcium release increases cytoplasmic and mitochondrial calcium, stimulating ROS production and lipid peroxidation.
- A positive feedback cycle involving IP3R-mediated calcium release contributes to ferroptosis execution.
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