The GRB10 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with disruption of the GRB10 gene in the NCI-H1975 human lung adenocarcinoma cell line. This product provides a genetically heterogeneous pool of knockout cells, avoiding clonal variation of single-cell-derived lines. CRISPR/Cas9-mediated gene disruption yields a mixed population with targeted modifications at the GRB10 locus, ideal for stable gene knockout studies with minimal selection bias in a cancer model.
The host cell line NCI-H1975 is a human lung adenocarcinoma epithelial cell line derived from lymph node metastasis. It harbors activating EGFR mutations (L858R and T790M) that confer constitutive kinase activity and resistance to first-generation tyrosine kinase inhibitors, making it a clinically relevant model for EGFR-driven non-small cell lung cancer (NSCLC) and drug resistance.
GRB10 encodes an adapter protein that negatively regulates insulin and IGF-1 receptor signaling. It binds phosphorylated INSR, IGF1R, and insulin receptor substrates IRS1/2, suppressing downstream PI3K/AKT and MAPK/ERK pathways. GRB10 interacts with NEDD4 to modulate receptor ubiquitination. Key effectors include AKT, ERK, and S6 kinase, resulting in reduced cell proliferation and growth. Thus, GRB10 acts as a brake on insulin/IGF-1 anabolic signaling.
In NCI-H1975 cells with hyperactive EGFR signaling due to L858R/T790M mutations, GRB10 disruption is predicted to relieve negative feedback and potentiate oncogenic PI3K/AKT and MAPK/ERK signaling. This may synergistically enhance proliferation, survival, and transformation. The model enables interrogation of cross-talk between insulin/IGF-1 and EGFR pathways, and investigation of how GRB10 modulates therapeutic responses to EGFR inhibitors. Researchers can dissect GRB10’s role in signal integration and identify potential vulnerabilities.
Applications include western blotting and phospho-signaling analysis of AKT, ERK, and S6 kinase upon insulin/IGF-1 stimulation. Proliferation and colony formation assays quantify growth effects, while drug sensitivity studies with EGFR inhibitors (e.g., gefitinib, osimertinib) reveal altered responses. The cells suit co-culture, xenograft models, and high-content screening for synthetic lethality or novel targets. This NSCLC model supports cancer biology, signal transduction, and precision medicine research. For additional details, contact Ascent Research.
