GPNMB Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for studying GPNMB gene function. The product comprises a heterogeneous pool of NCI-H1975 cells with targeted disruptions in GPNMB, generated via CRISPR/Cas9-mediated gene disruption. This polyclonal model avoids clonal biases and captures diverse editing outcomes, making it suitable for robust phenotypic analysis in cancer and cell biology research.
The parental NCI-H1975 line is a human lung adenocarcinoma cell line from a female nonsmoker, carrying EGFR T790M and L858R mutations. These mutations drive oncogenic signaling and confer resistance to first-generation tyrosine kinase inhibitors, establishing NCI-H1975 as a key model for EGFR-mutant non-small cell lung cancer (NSCLC) and a relevant system for investigating tumor cell migration and invasion.
GPNMB encodes a transmembrane glycoprotein that promotes cell adhesion and migration by binding integrins ??v??3 and ??v??5, leading to FAK phosphorylation and ERK1/2 and AKT activation. Its transcription is controlled by the MITF transcription factor and TGF-??, and its ectodomain is shed by ADAM10, yielding a soluble factor that suppresses T cell activation. GPNMB also interacts with syndecan-4 and upregulates MMP expression, linking it to tissue invasion and bone regulation. This signaling axis is critical for cellular motility and invasive programs.
In NCI-H1975 cells, GPNMB knockout allows dissection of integrin-mediated pathways that intersect with oncogenic EGFR signaling. Given GPNMB??s role in promoting migration and immune suppression, its disruption may impair metastatic and immunosuppressive phenotypes, offering insights into EGFR-mutant NSCLC progression. The polyclonal knockout population provides a robust system by averaging out clonal variation, supporting consistent results in functional screens.
Applications include wound-healing and transwell assays for migration/invasion, western blotting for phospho-FAK, -ERK1/2, and -AKT, flow cytometry for integrin surface expression, immunofluorescence of focal adhesions, co-immunoprecipitation of GPNMB-integrin complexes, and ADAM10 activity assays. These cells are valuable for studying cancer cell invasion, EGFR-mutant lung cancer biology, immune checkpoint modulation, bone metastasis, and Parkinson??s disease mechanisms. For further technical details or ordering information, please contact Ascent Research.
