The GSDME Knockout AGS Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population originating from the AGS human gastric epithelial adenocarcinoma line. This product provides a loss-of-function model for studying the tumor suppressor GSDME (DFNA5) through targeted gene disruption. As a polyclonal pool, it contains a heterogeneous mix of edited alleles, enabling robust bulk functional assays without clonal selection. Researchers can utilize these cells to probe GSDME-dependent pathways in a physiologically relevant gastric cancer environment.
The AGS parental line was established from a gastric adenocarcinoma of a human patient and is extensively employed as a model for gastric cancer biology. AGS cells display hallmark traits of gastric epithelial carcinoma, including rapid proliferation, anchorage-independent growth, and sensitivity to chemotherapeutic agents. Their well-characterized molecular background facilitates investigations into tumorigenesis, metastasis, and drug resistance. The introduction of a GSDME knockout into this system yields a controlled, isogenic platform for dissecting gene-specific functions relevant to gastric tumor biology.
GSDME functions as a critical executor of pyroptosis, a lytic and inflammatory cell death modality. Upon activation, caspase-3 cleaves GSDME, liberating its N-terminal fragment that oligomerizes to form membrane pores, resulting in cell lysis and the passive release of pro-inflammatory factors such as IL-1??, IL-18, HMGB1, and LDH. GSDME physically interacts with caspase-3, caspase-7, and the NLRP3 inflammasome component ASC, thereby bridging apoptotic signaling to pyroptotic execution. The gene is under transcriptional control of p53 and is frequently silenced by promoter DNA methylation in multiple cancer types. Its re-expression, triggered by chemotherapeutic agents, can redirect apoptosis toward immunogenic pyroptosis. Upstream death receptors (Fas, TRAIL-R) and TNF-?? converge on caspase-3 activation.
In the AGS gastric cancer context, GSDME is often epigenetically silenced, and its knockout recapitulates the loss-of-function state observed in many gastric tumors. This model enables researchers to investigate the impact of GSDME deficiency on chemosensitivity, inflammatory cytokine release, and tumor cell survival. The cells are particularly valuable for dissecting the cell death switch from apoptosis to pyroptosis induced by caspase-3-activating chemotherapeutics like cisplatin and doxorubicin. Comparative studies with parental AGS cells or GSDME-re-expressing counterparts help delineate the specific contribution of pyroptosis to tumor cell elimination and associated immune responses.
A diverse array of assays can be performed with these knockout cells, including LDH release and propidium iodide uptake to quantify cell lysis and membrane permeability, Western blotting to detect GSDME cleavage, and co-immunoprecipitation to analyze interactions with caspase-3. Transcript and epigenetic analyses may involve RT-qPCR and methylation-specific PCR, while cytokine secretion (IL-1??, IL-18) is measurable via ELISA. These tools facilitate investigations into pyroptosis in gastric cancer, epigenetic restoration of GSDME expression, and screening of therapeutic compounds that modulate inflammatory cell death. For additional technical information or custom application support, please contact Ascent Research.
