This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human hepatocellular carcinoma cell line Huh-7, engineered for targeted disruption of the CCDC50 gene. The polyclonal format preserves genetic heterogeneity, enabling robust loss-of-function studies without clonal selection bias. CRISPR/Cas9-mediated gene disruption ablates CCDC50 expression, providing a versatile model to dissect its biological roles in signal transduction and cellular homeostasis.
The parental Huh-7 cell line is a widely used model of hepatocellular carcinoma, originally established from a 57-year-old male patient. These cells exhibit epithelial morphology and retain key hepatic metabolic functions, making them particularly suitable for investigating liver cancer biology. Huh-7 cells harbor molecular features characteristic of advanced hepatocellular carcinoma, including dysregulated growth factor signaling and altered apoptotic pathways, thus offering a clinically relevant context for studying tumor suppressor mechanisms.
CCDC50 encodes an adaptor protein that functions as a negative regulator of pro-inflammatory NF-??B and oncogenic Wnt/??-catenin signaling. Mechanistically, CCDC50 is activated by inflammatory cytokines such as TNF-?? and interacts with A20/TNFAIP3 to recruit ubiquitin-editing machinery to its binding partner RIP1, promoting RIP1’s lysosomal degradation and thereby attenuating IKK complex-mediated NF-??B activation. In parallel, CCDC50 interacts with Dishevelled (DVL2) to inhibit ??-catenin stabilization downstream of LRP5/6, while its association with ATG16L1 facilitates LC3 lipidation and autophagosome formation. Through these interactions, CCDC50 acts as a molecular scaffold that coordinates endosomal sorting and autophagy, with downstream effects including RIP1 degradation, ??-catenin suppression, and enhanced autophagic flux.
In the context of Huh-7 hepatocellular carcinoma cells, CCDC50 loss is hypothesized to derepress NF-??B and Wnt pathways, promoting inflammatory and pro-survival signaling that may drive tumor progression. Given that chronic inflammation and autophagy dysfunction are hallmarks of hepatocellular carcinoma, this knockout model enables rigorous interrogation of CCDC50’s putative tumor suppressor role. The model is instrumental for dissecting how CCDC50-mediated cross-talk between autophagy and inflammatory signaling pathways impacts liver cancer cell proliferation, survival, and metastatic potential.
Researchers can employ this polyclonal knockout cell population in diverse applications, including western blotting to assess NF-??B (e.g., phospho-p65) and Wnt (e.g., active ??-catenin) pathway components, RT-qPCR profiling of downstream target genes, and autophagy flux assays monitoring LC3-II turnover. Functional studies may encompass cell proliferation, apoptosis, and migration/invasion experiments, as well as drug sensitivity screening to evaluate compounds targeting these pathways. Co-immunoprecipitation assays can further clarify CCDC50’s protein?Cprotein interactions. These applications facilitate detailed mechanistic studies and drug target validation programs. For additional technical details or order requests, please contact Ascent Research.
