CCDC50 Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human ccRCC cell line 786-O. This heterogeneous pool provides a loss-of-function model for studying CCDC50 without clonal artifacts, maintaining genetic diversity of the parental line. The cells enable investigation of CCDC50??s regulatory roles in cancer signaling and are suitable for functional genomics studies in ccRCC.
The parental 786-O line is a well-established human renal cell adenocarcinoma derived from a primary ccRCC of a 58-year-old male. It harbors a VHL truncating mutation leading to constitutive HIF pathway activation and is PTEN wild-type. The VHL loss synergizes with NF-kappaB signaling to promote oncogenic phenotypes, making this a clinically relevant model for ccRCC tumor biology and drug sensitivity studies.
CCDC50 encodes a ubiquitin-binding adaptor that negatively regulates NF-kappaB signaling by recruiting A20 to remove K63-linked ubiquitin chains from RIP1 and TRAF6, attenuating IKK and NF-kappaB activation. This reduces transcription of targets such as IL-6, TNF, and Bcl-xL, thereby dampening pro-inflammatory and survival signals. CCDC50 also modulates autophagy, evidenced by changes in LC3-II and p62 levels, and influences apoptotic pathways through caspase-3 cleavage. It interacts with the linear ubiquitin chain assembly complex (LUBAC) and IKK components, fine-tuning signal output. Upstream activation by TNF-alpha, LPS, IL-1beta, or CD40 ligand triggers CCDC50-dependent feedback inhibition, positioning it at the intersection of inflammation, cell survival, and death.
In VHL-mutant 786-O cells, CCDC50 disruption provides a powerful model to investigate NF-kappaB pathway dysregulation in ccRCC. VHL loss leads to constitutive HIF activation, which cooperates with NF-kappaB to drive proliferation and survival. Knockout of CCDC50 may enhance NF-kappaB activity, alter autophagic flux, and reduce apoptosis, enabling studies of tumor aggressiveness, metastatic potential, and drug resistance. This model is ideal for dissecting crosstalk between VHL/HIF and NF-kappaB signaling, as well as evaluating effects on cell migration and invasion.
Applications include Western blotting for NF-kappaB pathway components (phospho-IKK, IkB-alpha, p65) and autophagy markers (LC3-II, p62), NF-kappaB luciferase reporter assays, co-immunoprecipitation to assess ubiquitination, apoptosis detection via Annexin V/PI flow cytometry, and transwell migration/invasion assays. RNA sequencing can define CCDC50-dependent transcriptome changes. These cells are well-suited for screening modulators of inflammatory signaling and validating therapeutic targets in ccRCC. For additional details, contact Ascent Research.
