The CCL7 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the human lung adenocarcinoma line NCI-H1299, engineered to disrupt the CCL7 gene. The knockout is achieved through Cas9-mediated double-strand breaks and subsequent non-homologous end joining repair, generating a heterogeneous pool of CCL7-deficient cells. This polyclonal format avoids clonal selection artifacts and provides a robust loss-of-function model for studying the biological roles of CCL7 in cancer biology.
The NCI-H1299 cell line originates from a lymph node metastasis of a 43-year-old male patient with non-small cell lung adenocarcinoma. It exhibits characteristics of epithelial cells with high metastatic potential, rapid proliferation, and capacity for invasion. Widely used in lung cancer research, these cells lack functional p53 and serve as a model for aggressive tumor behavior. CCL7 disruption in this metastatic background allows examination of chemokine-mediated effects on tumor progression.
CCL7 (MCP-3) is a CC chemokine that acts as a chemoattractant for monocytes, eosinophils, and basophils via binding to G protein-coupled receptors CCR1, CCR2, and CCR3. Receptor engagement triggers intracellular signaling through MAPK/ERK and PI3K/AKT pathways, often involving PLC and G protein subunits. Expression of CCL7 is regulated by pro-inflammatory stimuli such as TNF, IL-1??, and IFN-??, acting through transcription factors NF-kB and STAT3. Downstream effects include upregulation of matrix metalloproteinases (MMPs) and vascular endothelial growth factor (VEGF). CCL7 also interacts with glycosaminoglycans to enhance local retention, amplifying its chemotactic activity.
In NCI-H1299 cells, CCL7 knockout disrupts autocrine and paracrine signaling loops that may influence cell migration, invasion, and proliferation. Loss of CCL7 can impair chemotactic responses and modify expression of MMPs and VEGF, potentially altering extracellular matrix remodeling and angiogenesis. This model helps elucidate how CCL7-mediated activation of ERK and AKT contributes to the metastatic phenotype of lung adenocarcinoma. The polyclonal nature captures functional heterogeneity relevant to tumor cell plasticity.
Research applications include quantitative gene expression analysis by RT-qPCR, protein detection via western blotting and ELISA, and functional assays such as transwell migration, invasion, and chemotaxis assays. Phospho-ERK and phospho-AKT analysis through flow cytometry or western blotting enables interrogation of downstream signaling. Additionally, immunofluorescence and cell viability assays support phenotypic characterization. These cells are suited for drug target validation and tumor microenvironment studies. For technical inquiries about this product and related services, please contact Ascent Research.
