The CCDC22 Knockout SK-HEP-1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of SK-HEP-1 human liver adenocarcinoma cells with targeted disruption of CCDC22. This heterogeneous pool circumvents clonal selection, providing a robust loss-of-function model that captures the variability of gene editing and enables pooled analysis of CCDC22-dependent phenotypes.
The SK-HEP-1 host cell line was derived from ascites of a male patient with liver adenocarcinoma and displays dual epithelial and endothelial features, making it a versatile model for hepatocellular carcinoma (HCC) studies. These adherent cells are tumorigenic and support investigations of tumor microenvironment interactions, metastasis, and angiogenic signaling, with extensive passaging capability suited for high-throughput applications.
CCDC22 is a subunit of the COMMD/CCDC22/CCDC93 (CCC) complex that recruits CUL2-RING ubiquitin ligase to mediate I-kappa-B alpha degradation, thereby activating NF-kappa-B signaling. Upon stimulation by TNF-alpha, IL-1beta, or TLR agonists, the IKK complex phosphorylates I-kappa-B alpha, marking it for CCC/CUL2-dependent ubiquitination and proteasomal turnover, which liberates NF-kappa-B p50/p65 dimers for nuclear translocation and target gene induction. CCDC22 interacts with COMMD1, CCDC93, and other COMMD family proteins, positioning it as a positive regulator of NF-kappa-B-driven transcription of pro-inflammatory cytokines (IL6, TNF, CXCL8) and anti-apoptotic factors like BCL2.
In SK-HEP-1 HCC cells, constitutive or inducible NF-kappa-B activity contributes to inflammation-associated oncogenesis, chemoresistance, and immune evasion. CCDC22 disruption impairs stimulus-coupled I-kappa-B degradation, dampening canonical NF-kappa-B transcriptional responses and enabling dissection of pathway contributions to proliferation, apoptosis, and the tumor secretome. This model is relevant for studying Ritscher-Schinzel syndrome, immune disorders, and inflammation-driven cancers.
Applications include western blot analysis of IKK/I-kappa-B/NF-kappa-B phosphorylation and degradation, RT-qPCR profiling of NF-kappa-B target genes, and NF-kappa-B luciferase reporter assays to measure transcriptional activity. Co-immunoprecipitation experiments assess CCC complex integrity without CCDC22, while proliferation, apoptosis, and multiplex cytokine secretion assays reveal functional consequences. These tools facilitate exploration of NF-kappa-B biology in HCC and screening of pathway modulators. For inquiries, contact Ascent Research.
