The CCDC50 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 non-small cell lung adenocarcinoma cell line. This product contains a heterogeneous mixture of cells with targeted disruption of the CCDC50 gene, providing a robust loss-of-function model for studying CCDC50-mediated regulatory mechanisms. The polyclonal format captures the diversity of editing outcomes, enabling assessment of functional consequences without clonal selection bias. These cells are designed for advanced research applications requiring abrogation of CCDC50 expression to dissect its role in signal transduction and cellular homeostasis.
NCI-H1975 is a well-characterized human lung adenocarcinoma epithelial cell line harboring an activating EGFR exon 19 deletion and a TP53 missense mutation, recapitulating key genetic features of aggressive NSCLC. These adherent cells are widely used as a model system for lung cancer biology, drug response, and resistance mechanisms. The line’s dependency on EGFR signaling and p53 deficiency make it particularly valuable for investigating interplay between oncogenic drivers and tumor suppressor pathways. Integrating the CCDC50 knockout into this genetic background allows researchers to explore how autophagy-mediated regulation of NF-kB intersects with established oncogenic signaling networks in a clinically relevant context.
CCDC50 functions as a negative regulator of Toll-like receptor (TLR)-mediated NF-kB signaling. Upon TLR3 activation by poly(I:C) or TLR4 stimulation by LPS, downstream adaptor MyD88 triggers K63-linked polyubiquitination of NEMO (IKBKG), a critical step for NF-kB transcriptional activity. CCDC50 specifically recognizes K63-polyubiquitinated NEMO and targets it for autophagic degradation via interaction with the autophagy receptor SQSTM1/p62 and the autophagosomal marker MAP1LC3B (LC3B). This selective autophagy of activated NEMO limits NF-kB activation and subsequent transcription of pro-inflammatory target genes including IL6 and TNF. Consequently, CCDC50 serves as a molecular brake on innate immune signaling, linking ubiquitin-dependent protein quality control to modulation of inflammatory gene expression.
In NCI-H1975 cells, constitutive EGFR signaling and p53 loss create a pro-inflammatory environment. CCDC50 knockout likely relieves NF-kB inhibition, enhancing transcription of pro-survival factors, altering proliferation and apoptosis sensitivity, and disrupting autophagy dynamics. Since autophagy can be tumor-suppressive or tumor-promoting, loss of CCDC50-mediated selective degradation may shift cellular responses to EGFR inhibitors. This polyclonal knockout population is a versatile tool to examine how deregulated NF-kB and autophagy contribute to adenocarcinoma malignancy.
This knockout model supports multifaceted investigation of CCDC50 function. Key applications include western blotting of NF-kB pathway components (p65, I??B??) and autophagy markers (LC3B, SQSTM1/p62), RT-qPCR of NF-kB targets (IL6, TNF), luciferase reporter assays, co-immunoprecipitation of CCDC50-NEMO, and autophagy flux analysis. Functional studies such as proliferation, apoptosis, migration/invasion, and drug sensitivity screening can reveal phenotypic consequences. This system is ideal for dissecting innate immunity modulation and therapeutic resistance in lung adenocarcinoma. For technical inquiries or a quote, contact Ascent Research.
