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Published on: May 31, 2024
Detection of Gasdermin C-Mediated Cancer Cell Pyroptosis
Junwei Hou1,2,3,4,5,6, Shuanglian Wang3,4, Rui Miao1,3,4
1Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital, Central South University, Changsha, Hunan, China.
Abstract:
Gasdermin proteins except DFNB59 are the executioners of pyroptotic cell death. Cleavage of a gasdermin by an active protease causes lytic cell death. Gasdermin C (GSDMC) is cleaved by caspase-8 in response to macrophage-secreted TNF-α. Upon cleavage, the GSDMC-N domain is liberated and oligomerized, followed by pore formation in the plasma membrane. GSDMC cleavage, LDH release, and plasma membrane translocation of GSDMC-N domain are the reliable markers for GSDMC-mediated cancer cell pyroptosis (CCP). Here, we describe the methods for examining GSDMC-mediated CCP.
Insights
Gasdermin C (GSDMC) triggers pyroptosis in cancer cells. This study details methods to detect GSDMC-mediated cancer cell pyroptosis (CCP) using key molecular and cellular markers.
Area of Science:
- Molecular Biology
- Cell Death Research
- Cancer Biology
Background:
- Gasdermin proteins are key mediators of pyroptosis, a programmed lytic cell death.
- Gasdermin C (GSDMC) is activated by caspase-8 cleavage, induced by tumor necrosis factor-alpha (TNF-α).
- GSDMC activation leads to N-terminal domain liberation, oligomerization, and plasma membrane pore formation.
Purpose of the Study:
- To describe methodologies for the detection and analysis of GSDMC-mediated cancer cell pyroptosis (CCP).
- To establish reliable markers for studying GSDMC's role in cancer cell death.
Main Methods:
- Assessing GSDMC cleavage by specific proteases.
- Quantifying lactate dehydrogenase (LDH) release as a marker of cell lysis.
- Analyzing the translocation of the GSDMC-N terminal domain to the plasma membrane.
Main Results:
- GSDMC cleavage, LDH release, and GSDMC-N domain plasma membrane translocation are validated as reliable indicators of CCP.
- These markers provide a robust framework for investigating GSDMC's function in cancer cell pyroptosis.
Conclusions:
- The described methods enable accurate examination of GSDMC-mediated cancer cell pyroptosis.
- This research facilitates further understanding of pyroptosis pathways in cancer therapeutics.
