The GPRC5A Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population featuring targeted disruption of the GPRC5A tumor suppressor gene in human NCI-H1975 lung adenocarcinoma epithelial cells. This loss-of-function model enables detailed investigation of GPRC5A-mediated signaling within an EGFR-driven oncogenic context, providing a genetically diverse system that avoids clonal selection artifacts.
The NCI-H1975 host cell line is derived from non-small cell lung carcinoma and harbors the EGFR double mutant L858R/T790M, conferring constitutive kinase activity and resistance to first-generation tyrosine kinase inhibitors. These adherent epithelial cells are widely used as a tumorigenic model to study EGFR-dependent oncogenesis, acquired drug resistance, and molecular mechanisms of lung adenocarcinoma progression, making them highly relevant for preclinical therapeutic evaluation.
GPRC5A encodes a retinoic acid-inducible orphan G protein-coupled receptor that functions as a tumor suppressor. It is transcriptionally regulated by retinoic acid-activated RAR/RXR heterodimers, p53, and SP1. Mechanistically, GPRC5A inhibits EGFR phosphorylation, thereby attenuating downstream STAT3 and NF-??B activation, which leads to downregulation of cyclin D1, destabilization of ??-catenin, and activation of caspase-3-mediated apoptosis. Additionally, GPRC5A interacts with TRAF2 and G?? proteins, further integrating retinoic acid responses with oncogenic signaling pathways.
In the EGFR-hyperactive NCI-H1975 background, ablation of GPRC5A disrupts a critical negative regulatory node, resulting in enhanced EGFR/STAT3/NF-??B signaling and increased tumorigenic potential. This polyclonal knockout model recreates aspects of intratumoral heterogeneity, allowing researchers to dissect the cooperative effects of GPRC5A loss and EGFR mutations on lung adenocarcinoma cell proliferation, survival, and drug resistance.
These cells are suitable for a broad range of applications, including dissection of EGFR and retinoic acid signaling networks, functional analysis of tumor suppressor genes, and profiling of therapeutic sensitivities. Representative assays include Western blotting for phosphorylated EGFR, STAT3, and NF-??B; RT-qPCR for cyclin D1 and caspase-3; cell proliferation (MTT); apoptosis (Annexin V); migration and invasion (Transwell); and dose?Cresponse studies with EGFR inhibitors such as osimertinib. The polyclonal knockout pool also supports synthetic lethal screening. For technical inquiries or ordering, please contact Ascent Research.
