The GSDMD Knockout NCI-H1975 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma cell line. This polyclonal knockout model provides a loss-of-function system for studying gasdermin D (GSDMD) function in a non-small cell lung carcinoma (NSCLC) background. The polyclonal nature of the knockout population ensures representation of diverse GSDMD-disrupted genotypes, avoiding clonal selection artifacts. This product is designed for researchers investigating pyroptosis, inflammasome signaling, and inflammatory cell death mechanisms.
The NCI-H1975 host cell line is a well-characterized model of lung adenocarcinoma, isolated from a human female. These cells harbor EGFR L858R/T790M mutations, conferring resistance to first- and second-generation EGFR tyrosine kinase inhibitors (TKIs) such as gefitinib. NCI-H1975 cells are tumorigenic and widely used to study mechanisms of TKI resistance, oncogenic signaling, and therapeutic responses in NSCLC. Their epithelial origin and retention of key lung cancer pathways make them a suitable platform for interrogating the role of GSDMD in inflammation-associated cancer phenotypes.
GSDMD encodes a pore-forming protein that acts as the central executioner of pyroptotic cell death. Upon activation of canonical or non-canonical inflammasomes, inflammatory caspases??including caspase-1, caspase-4, caspase-5, and caspase-11??cleave GSDMD, releasing its N-terminal domain. This fragment oligomerizes and inserts into the plasma membrane to form pores, leading to cell lysis, release of pro-inflammatory cytokines such as IL-1?? and IL-18, and propagation of inflammation. Upstream regulators include NLRP3, AIM2, and NLRC4 inflammasomes, type I interferons, and LPS. Key downstream events include HMGB1 secretion, LDH release, and inflammatory cytokine secretion. GSDMD function is linked to ASC, NINJ1, and the TLR4/NF-??B axis, positioning it at the nexus of immune detection and inflammatory cell death.
In the context of EGFR-mutant lung adenocarcinoma, GSDMD-mediated pyroptosis may contribute to tumor inflammation, immune cell recruitment, and drug resistance. NCI-H1975 cells with EGFR T790M mutation represent a clinically relevant resistance model, and GSDMD disruption can help elucidate whether pyroptosis influences TKI sensitivity or the tumor microenvironment. This polyclonal knockout model allows dissection of GSDMD-dependent pathways in a background that mimics aggressive NSCLC, providing insights into potential therapeutic vulnerabilities where modulating inflammatory cell death could enhance anti-tumor immunity or overcome resistance.
Researchers can employ this polyclonal knockout population in various functional assays. Western blotting confirms loss of full-length GSDMD and its cleaved fragment upon inflammasome stimulation. Pyroptosis induction with LPS/nigericin, followed by LDH release and IL-1??/IL-18 ELISA, quantifies pore formation and cytokine release. Flow cytometry for propidium iodide uptake monitors membrane integrity loss. Caspase-1 activation and cell viability assays further characterize inflammasome engagement. EGFR inhibitor sensitivity testing can explore GSDMD?CTKI response interplay. For further technical information or orders, contact Ascent Research.
