Suppression of CLIC4/mtCLIC enhances hydrogen peroxide-induced apoptosis in C6 glioma cells
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Updated: May 14, 2026
Detection of Mitochondria Membrane Potential to Study CLIC4 Knockdown-induced HN4 Cell Apoptosis In Vitro
Published on: July 17, 2018
Ye Xu1, Jinsong Kang, Zhaoxin Yuan
1Department of Pathophysiology, Norman Bethune College of Medicine, Jilin University, Changchun 130021, PR China.
Abstract:
CLIC4/mtCLIC (referred to here as CLIC4) is one of the seven-member family of chloride intracellular channels (CLIC). CLIC4 localizes to the mitochondria, nucleus, cytoplasm and other organellular compartments and participates in the apoptotic response to stress. However, the role of CLIC4 in oxidative stress and apoptosis is not well understood. In this study, we showed the important role of CLIC4 in apoptosis of C6 glioma cells induced by hydrogen peroxide (H2O2). Our results showed that CLIC4 protein expression was upregulated following H2O2-induced C6 cell apoptosis. The upregulation of CLIC4 protein expression was paralleled with an increased Bax/Bcl-2 ratio, cytochrome c and cleaved caspase-3 protein expression upon H2O2-induced C6 cell apoptosis. Suppression of CLIC4 expression by RNA interference enhanced cell apoptosis, but the ratio of Bax/Bcl-2 was not involved in this process. Dissipation of mitochondrial membrane potential and nuclear translocation of CLIC4 were involved in the activation of apoptosis induced by H2O2. Our data indicate that CLIC4 protein may be a key element in the apoptotic response to oxidative stress.
Insights
Chloride intracellular channel 4 (CLIC4) plays a key role in hydrogen peroxide-induced apoptosis in C6 glioma cells. Its upregulation and nuclear translocation are critical for oxidative stress-induced cell death.
Area of Science:
- Cell Biology
- Molecular Biology
- Neuroscience
Background:
- Chloride intracellular channels (CLIC) are a family of proteins involved in cellular processes.
- CLIC4 localizes to various cellular compartments and participates in stress-induced apoptosis.
- The specific role of CLIC4 in oxidative stress and apoptosis remains unclear.
Purpose of the Study:
- To investigate the role of CLIC4 in hydrogen peroxide (H2O2)-induced apoptosis in C6 glioma cells.
- To elucidate the molecular mechanisms by which CLIC4 influences oxidative stress response.
Main Methods:
- Utilized C6 glioma cells and induced apoptosis with hydrogen peroxide (H2O2).
- Assessed CLIC4 protein expression, Bax/Bcl-2 ratio, cytochrome c, and cleaved caspase-3.
- Employed RNA interference to suppress CLIC4 expression.
- Monitored mitochondrial membrane potential and CLIC4 nuclear translocation.
Main Results:
- CLIC4 protein expression was upregulated in H2O2-induced C6 cell apoptosis.
- Increased Bax/Bcl-2 ratio, cytochrome c, and cleaved caspase-3 accompanied CLIC4 upregulation.
- CLIC4 suppression enhanced apoptosis, independent of the Bax/Bcl-2 ratio.
- Dissipation of mitochondrial membrane potential and CLIC4 nuclear translocation were observed.
Conclusions:
- CLIC4 is significantly involved in the apoptotic response of C6 glioma cells to oxidative stress.
- CLIC4 protein may serve as a key mediator in apoptosis triggered by H2O2.
- Nuclear translocation of CLIC4 and mitochondrial membrane potential changes are integral to H2O2-induced apoptosis activation.