The CCDC50 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population derived from the SK-HEP-1 human hepatic adenocarcinoma cell line, in which the coiled-coil domain-containing protein 50 (CCDC50) gene has been disrupted. This product provides a loss-of-function model for investigating CCDC50-dependent autophagy, epidermal growth factor receptor (EGFR) signaling, and NF-kappaB activation in a liver cancer context. The polyclonal nature ensures a heterogeneous mix of edited alleles, enabling the study of gene disruption effects without clonal selection artifacts.
The SK-HEP-1 host cell line originates from ascites of a patient with liver adenocarcinoma and exhibits a hybrid phenotype combining epithelial and endothelial characteristics, often used as a surrogate for liver sinusoidal endothelial cells. These cells are widely employed in hepatocellular carcinoma research, metastasis studies, and drug metabolism assays. Their ability to form tumors and recapitulate aspects of the liver microenvironment makes them a relevant platform for dissecting molecular mechanisms underlying hepatic carcinogenesis.
CCDC50 serves as a multifunctional adaptor protein that links extracellular signals to autophagy and NF-kappaB pathways. Upon EGF stimulation, CCDC50 interacts with EGFR, Beclin1, and LC3 to promote autophagosome formation while simultaneously preventing EGFR degradation, thereby sustaining downstream PI3K/AKT/mTOR signaling. Additionally, CCDC50 modulates NF-kappaB activation through interaction with the IKK complex and regulates apoptosis by influencing caspase activity. Cellular stress stimuli and TNF-alpha also engage CCDC50, highlighting its role as a signaling hub integrating autophagy, survival, and inflammatory responses.
In SK-HEP-1 cells, CCDC50 knockout disrupts the balance between EGFR-driven proliferation and autophagic degradation, potentially sensitizing these liver cancer cells to EGFR-targeted therapies or autophagy modulators. The loss of CCDC50 may alter NF-kappaB-mediated transcription of pro-survival genes and affect apoptosis sensitivity, providing a model to explore resistance mechanisms in hepatocellular carcinoma. This system allows researchers to dissect the interplay between autophagy and oncogenic signaling in a cell type that retains both epithelial and endothelial features, reflecting the complexity of the tumor microenvironment.
Researchers can employ this knockout model for a range of applications, including screening for autophagy modulators, assessing EGFR degradation kinetics via Western blot or cycloheximide chase assays, and quantifying NF-kappaB activity using luciferase reporters. Co-immunoprecipitation experiments can validate CCDC50’s interaction with Beclin1 or SQSTM1/p62, while immunofluorescence allows visualization of LC3 puncta as a measure of autophagic flux. Apoptosis assays by flow cytometry further enable studies on cell death pathways. For additional details on this product, contact Ascent Research.
