The GOLPH3 Knockout NCI-H1975 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population with disrupted GOLPH3 in the NCI-H1975 human lung adenocarcinoma cell line. This heterogeneous loss-of-function model captures multiple editing events, minimizing clonal bias and enabling pooled functional screens, signaling analyses, and drug response studies in the context of EGFR-mutant non-small cell lung cancer (NSCLC).
NCI-H1975 is a female-derived metastatic pleural effusion cell line carrying the oncogenic EGFR L858R mutation and a TP53 mutation, widely applied in EGFR signaling and tyrosine kinase inhibitor resistance research. The L858R mutation constitutively activates EGFR and downstream PI3K/AKT and MAPK cascades, driving tumor cell proliferation and survival.
GOLPH3 is a Golgi-resident oncoprotein that interacts with MYO18A to regulate Golgi morphology and vesicular trafficking. It functions upstream of the PI3K/AKT/mTOR pathway by enhancing glycosylation-dependent stabilization of growth factor receptors, thereby activating AKT and mTOR. GOLPH3 is regulated by MYC, growth factors, and ER stress, and its downstream effectors include AKT, mTOR, MMP9, cyclin D1, and BCL2. Additional interactions with GOLGA2, GORASP2, and DNA-PKcs link GOLPH3 to Golgi architecture maintenance and DNA damage repair. Thus, GOLPH3 integrates Golgi trafficking, signal transduction, and cell survival pathways.
In NCI-H1975 cells, GOLPH3 knockout disrupts a critical node coupling Golgi function to oncogenic EGFR signaling. Since EGFR L858R requires proper glycosylation and surface localization, loss of GOLPH3 likely impairs EGFR stabilization and downstream AKT/mTOR output, making this model valuable for elucidating GOLPH3-dependent EGFR modulation, drug resistance mechanisms, and the Golgi??s role in metastasis. The inherent resistance of NCI-H1975 to some EGFR inhibitors further positions the knockout pool as a platform for studying adaptive signaling networks.
Typical applications include western blotting for GOLPH3 and phospho-AKT, immunofluorescence for Golgi markers, transwell migration/invasion assays, cell proliferation assays, and drug sensitivity testing with EGFR inhibitors. Co-immunoprecipitation with MYO18A confirms protein interactions, and RNA-seq reveals transcriptome-wide impact. This knockout model supports investigation of GOLPH3-mediated EGFR signaling, Golgi trafficking in cancer, drug resistance in EGFR-mutant NSCLC, and metastasis dependency on Golgi function. For further information or custom gene-editing inquiries, please contact Ascent Research.
