The INSR Knockout NCI-H1975 Polyclonal Cells comprise a heterogeneous population of CRISPR/Cas9-edited NCI-H1975 cells bearing targeted disruption of the human INSR locus. This polyclonal knockout cell product provides a loss-of-function model for studying insulin receptor signaling without the need for single-cell clonal isolation. By ablating INSR expression, researchers gain a genetically defined tool to dissect receptor tyrosine kinase-dependent mechanisms in a lung adenocarcinoma background.
The parental NCI-H1975 cell line was originally derived from the pleural effusion of a female patient with nonsmall cell lung cancer (NSCLC), adenocarcinoma subtype. These epithelial cells harbor an activating mutation in the epidermal growth factor receptor (EGFR L858R) and a secondary T790M gatekeeper mutation, rendering them a clinically relevant model for EGFR-mutant NSCLC. Their cancerous properties, including dysregulated proliferation and metabolic adaptation, make them an appropriate host for interrogating the interplay between oncogenic drivers and insulin receptor function.
The INSR gene encodes the insulin receptor tyrosine kinase, activated by insulin, IGF-1, and IGF-2. Upon binding, it autophosphorylates and recruits IRS1, SHC, and GRB2, engaging the PI3K-Akt-mTOR and RAS-MAPK cascades. These pathways regulate glucose metabolism, cell growth, and survival via effectors like Akt, mTOR, ERK1/2, FOXO1, and GLUT4. Negative feedback is mediated by PTP1B and SOCS proteins.
Disruption of INSR in NCI-H1975 cells abolishes insulin/IGF-dependent activation of Akt and ERK, thereby impairing proliferative and survival signals. Given that NSCLC cells often rely on metabolic reprogramming and growth factor inputs, this knockout model enables dissection of insulin-mediated contributions to tumor malignancy. The polyclonal nature preserves the genetic diversity of the edited population, reflecting heterogeneous responses and offering a more robust representation of the in vivo scenario. Consequently, it is a valuable system for studying how loss of insulin signaling impacts cancer cell behavior and drug responses.
This INSR knockout cell product supports assays such as western blotting for phospho-Akt and phospho-ERK, RT-qPCR for INSR and IRS1, and phospho-RTK arrays. Metabolic readouts include glucose uptake and lactate production. Proliferation, apoptosis (annexin V), migration, and invasion assays can elucidate functional consequences. RNA-seq and co-immunoprecipitation further characterize the signaling landscape. Drug sensitivity testing with insulin analogs, metformin, or EGFR inhibitors can identify therapeutic targets. These cells are well-suited for studying insulin resistance, metabolic regulation in NSCLC, and resistance to EGFR inhibitors. For additional information, please contact Ascent Research.
