The CCDC50 Knockout A2780 Polyclonal Cells product provides a heterogeneous population of A2780 cells subjected to CRISPR/Cas9-mediated disruption of the CCDC50 gene, generating a loss-of-function model for studying the multifunctional adaptor protein CCDC50. As a polyclonal knockout pool, these cells offer a diverse genetic background that resembles population-level variability, making them suitable for functional screens and pathway analyses without clonal selection artifacts. This product is intended for advanced biomedical research, enabling interrogation of CCDC50-dependent signaling and autophagy mechanisms within an epithelial ovarian carcinoma context.
The parental A2780 cell line was established from an untreated patient with ovarian endometrioid adenocarcinoma and serves as a well-characterized model for ovarian cancer. These epithelial ovarian carcinoma cells retain key signaling features relevant to tumor progression, metastasis, and therapeutic response. By comparing CCDC50 knockout A2780 polyclonal cells to wild-type controls, researchers can assess the impact of CCDC50 disruption on ovarian cancer pathways, including NF-??B-driven inflammation and autophagy-associated survival.
CCDC50 is a cytoplasmic adaptor protein that acts as a negative regulator of NF-??B signaling by inhibiting ubiquitination of the receptor-interacting protein kinase RIP1, thereby attenuating the I??B kinase (IKK) complex activation downstream of TNFR1 and TLR stimulation. Under TNF-?? or IL-1?? stimulation, CCDC50 interacts with RIP1 and the regulatory subunit IKK??/NEMO, suppressing phosphorylation and degradation of I??B?? and subsequent nuclear translocation of NF-??B. Consequently, expression of NF-??B target genes such as IL6, IL8, and BCL2 is reduced. In addition, CCDC50 promotes autophagy by facilitating the association of signaling complexes with LC3 and the cargo receptor p62/SQSTM1, leading to lysosomal degradation of the TNFR1 complex and other ubiquitinated aggregates. This dual function positions CCDC50 as a key node connecting inflammatory responses and autophagic clearance.
In the A2780 ovarian cancer model, CCDC50 knockout enables investigation of how NF-??B hyperactivity and altered autophagy influence tumor cell proliferation, apoptosis, and drug resistance. Ovarian carcinomas often exhibit constitutive NF-??B activation and dysregulated autophagy, both of which contribute to chemoresistance and tumor maintenance. By disrupting CCDC50, this model can reveal mechanisms by which cancer cells exploit these pathways, offering a platform for testing interventions that target NF-??B or autophagy. Additionally, since CCDC50 mutations are associated with autosomal dominant nonsyndromic hearing loss (DFNA44), these cells may also be used to explore cell-type-specific functions of CCDC50 beyond cancer.
Researchers can employ this knockout model in a range of assays to probe CCDC50 function. Western blotting for NF-??B pathway components (phospho-I??B??, p65) and autophagy markers (LC3-II, p62) permits biochemical validation of pathway activation. NF-??B luciferase reporter assays quantify transcriptional activity, while co-immunoprecipitation confirms interactions with RIP1 or NEMO. Immunofluorescence detection of LC3 puncta and RT-qPCR for IL6 and IL8 expression provide functional readouts of autophagy and inflammation, respectively. Cell viability and apoptosis assays using chemotherapeutics can assess the contribution of CCDC50 to drug response. For further details on this product, please contact Ascent Research.
