The EGFL7 Knockout NCI-H1975 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population derived from the NCI-H1975 lung adenocarcinoma cell line, with targeted disruption of the EGFL7 gene. This loss-of-function model provides a heterogeneous cell pool for investigating EGFL7 roles in angiogenesis and Notch modulation without requiring clonal isolation.
NCI-H1975 is a human non-small cell lung cancer line harboring EGFR L858R and T790M mutations, driving constitutive kinase activity and resistance to first-generation EGFR-TKIs. It serves as a widely used model for EGFR-driven oncogenesis and therapeutic resistance in lung adenocarcinoma.
EGFL7 encodes a secreted protein that binds integrin ??v??3 (ITGAV/ITGB3) and activates focal adhesion kinase (FAK), leading to downstream phosphorylation of AKT and ERK, which promote endothelial cell migration and angiogenesis. In parallel, EGFL7 antagonizes DLL4-mediated Notch signaling by directly interacting with NOTCH1 and NOTCH4 receptors, thereby repressing transcription of canonical Notch target genes HES1 and HEY1. This dual function positions EGFL7 as a key node integrating endothelial adhesion and signaling. Its expression is transcriptionally regulated by the ETS factors ERG and FLI1, and by hypoxia-inducible HIF1A.
In the EGFR-mutant NCI-H1975 background, EGFL7 knockout provides a unique platform to dissect how this factor contributes to lung adenocarcinoma phenotypes. Although EGFL7 is predominantly characterized in endothelial cells, its potential expression in cancer cells may modulate integrin-mediated adhesion and signaling crosstalk with oncogenic EGFR pathways, influencing processes such as tumor angiogenesis, metastatic spread, and response to EGFR-TKIs like osimertinib. This model thus facilitates exploration of EGFL7??s role in NSCLC progression and drug resistance.
Detailed research applications include functional assays such as transwell migration and endothelial tube formation to assess angiogenic and motility changes upon EGFL7 disruption. Notch reporter assays and quantitative analysis of downstream targets (HES1, HEY1) reveal pathway activity. Biochemical validation employs Western blotting for EGFL7 protein, RT-qPCR for mRNA, and Sanger sequencing for indel detection. Phospho-signaling analyses probe FAK, AKT, and ERK activation status. Co-immunoprecipitation further characterizes EGFL7 interactions with ITGAV/ITGB3 and Notch receptors. Additionally, drug sensitivity testing with osimertinib can uncover EGFL7-dependent modulation of EGFR-TKI response in this resistant model. For further information, please contact Ascent Research.
