The GOLPH3L Knockout NCI-H1975 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human GOLPH3L gene in the NCI-H1975 lung adenocarcinoma cell line. This polyclonal format, generated by CRISPR/Cas9-mediated gene disruption, delivers a heterogeneous pool of edited cells that averages clonal variation, enabling robust loss-of-function studies in pooled screening and functional genomic applications. The cell population is designed to support research into Golgi biology, mTOR signaling, and the molecular underpinnings of EGFR-mutant non-small cell lung cancer (NSCLC).
The host cell line NCI-H1975 is a widely utilized human NSCLC model derived from a non-smoking female patient. It harbors activating EGFR L858R and T790M mutations, resulting in constitutive kinase activity and resistance to first-generation EGFR tyrosine kinase inhibitors (TKIs). These mutations drive persistent PI3K-AKT and MAPK pathway activation, rendering the cell line dependent on oncogenic signals essential for tumor maintenance, proliferation, and acquired drug resistance. Consequently, NCI-H1975 is a clinically relevant system for investigating mechanisms of TKI resistance and evaluating next-generation therapeutic strategies.
GOLPH3L encodes a Golgi-localized phosphoprotein that binds phosphatidylinositol-4-phosphate (PtdIns(4)P) and plays a central role in maintaining Golgi morphology and regulating vesicular trafficking. Functionally, GOLPH3L is integrated into the PI3K-AKT-mTOR signaling axis: it is activated by PtdIns(4)P and AKT and acts upstream of mTORC1, directly controlling the phosphorylation of its effector S6K1. GOLPH3L interacts with MYO18A and the COPI complex, physically linking Golgi structure to secretory pathway dynamics. Through these interactions, GOLPH3L couples nutrient-sensing and growth factor signals to Golgi architecture and cell growth, ultimately modulating mTORC1-mediated phosphorylation of S6K1 and 4E-BP1.
In the NCI-H1975 background, oncogenic EGFR signaling constitutively activates the PI3K-AKT pathway, which may converge on GOLPH3L at the Golgi to amplify mTORC1 activity and sustain malignant phenotypes. Disrupting GOLPH3L in this context allows dissection of its contribution to linking Golgi function with tumor cell proliferation, survival, and drug sensitivity. Moreover, GOLPH3L-dependent vesicular trafficking may influence the subcellular distribution and signaling output of EGFR, potentially affecting TKI response. Therefore, this knockout model enables interrogation of how GOLPH3L-driven Golgi dynamics and mTOR activation modulate resistance to EGFR inhibitors, including the third-generation TKI osimertinib, offering insights into novel combination therapies.
This polyclonal knockout population is compatible with a broad range of experimental approaches. GOLPH3L protein loss can be confirmed by Western blotting, while immunofluorescence and Golgi morphology analysis allow visualization of structural changes in the Golgi apparatus. mTOR signaling status can be assessed through phosphorylation-specific Western blots for S6K1 or 4E-BP1. Functional consequences of GOLPH3L disruption are measured using cell proliferation and migration assays, and drug sensitivity profiling??including to osimertinib??reveals impacts on therapeutic response. The product thus provides a versatile resource for cancer biology and signaling studies. For further details, please contact Ascent Research.
