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Updated: Jun 18, 2026
Evaluation of Caspase Activation to Assess Innate Immune Cell Death
Published on: January 20, 2023
Role of caspase 8 as a determinant in trail sensitivity of neuroblastoma cell lines
Haixia Tong1, Chunwei Lu, Yanmin Yang
1Laboratory of Hematology, Shengjing Hospital of China Medical University, Shenyang, China. tonghaixia18@yahoo.com.cn
Objective:
Resistance of neuroblastoma (NB) cells to tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-mediated apoptosis is thought to be caused by loss of caspase 8 expression. The aim of this study is to investigate if induction of caspase 8 by gamma-interferon (IFN gamma) renders NB cells sensitive to TRAIL.
Methods:
Caspase 8 expression was monitored by RT-PCR and Western blot analysis. The effects of IFN gamma, TRAIL, IFN gamma + TRAIL, and caspase 8 inhibitor + TRAIL on the growth and apoptosis of NB cells were detected with the methods of reduction rate of Alamar blue assay and flow cytometry. The relative caspase 8 activity was measured with colorimetric assay.
Results:
An increased expression of caspase 8 mRNA and protein was found after treatment with IFN gamma. IFN gamma in combination with TRAIL decreased proliferation of NB cell line SH-SY5Y cells. The killing effect of TRAIL on NB cells expressing caspase 8 was depressed by caspase 8 inhibitor. The relative caspase 8 activity of NB cells expressing caspase 8 increased with the prolongation of TRAIL action time.
Conclusions:
TRAIL induced apoptosis in NB cells expressing caspase 8.
Insights
Gamma-interferon (IFN gamma) induction of caspase 8 restores TRAIL sensitivity in neuroblastoma cells. This finding suggests a potential therapeutic strategy for neuroblastoma treatment.
Area of Science:
- Oncology
- Molecular Biology
- Cell Death Research
Background:
- Neuroblastoma (NB) cells often exhibit resistance to TRAIL-induced apoptosis.
- Loss of caspase 8 expression is a key mechanism underlying this TRAIL resistance.
Purpose of the Study:
- To investigate if gamma-interferon (IFN gamma) can induce caspase 8 expression in NB cells.
- To determine if IFN gamma-induced caspase 8 expression sensitizes NB cells to TRAIL-mediated apoptosis.
Main Methods:
- Caspase 8 expression analyzed via RT-PCR and Western blot.
- Cell proliferation and apoptosis assessed using Alamar blue assay and flow cytometry.
- Caspase 8 activity measured by colorimetric assay.
Main Results:
- IFN gamma treatment significantly increased caspase 8 mRNA and protein levels.
- Combined IFN gamma and TRAIL treatment reduced proliferation of SH-SY5Y NB cells.
- Caspase 8 inhibitor attenuated TRAIL's killing effect, confirming caspase 8's role.
- Increased caspase 8 activity correlated with prolonged TRAIL exposure time.
Conclusions:
- TRAIL effectively induces apoptosis in neuroblastoma cells that express caspase 8.
- IFN gamma can restore TRAIL sensitivity in NB cells by upregulating caspase 8.
