The CCDC85C Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human cell line, designed to disrupt the expression of the CCDC85C gene. This loss-of-function model provides a robust tool for dissecting the tumor-suppressive functions of CCDC85C within a liver adenocarcinoma background. The polyclonal nature of the knockout pool ensures a heterogeneous population that captures diverse editing events, enabling the study of gene disruption effects across a broad cellular context without clonal bias.
The SK-HEP-1 host cell line originates from the ascitic fluid of a patient with liver adenocarcinoma and was historically misclassified as hepatocellular carcinoma. It exhibits a mixed epithelial and mesenchymal phenotype, which contributes to its utility as a model for metastatic progression and epithelial-mesenchymal plasticity. Although subsequent characterization has identified an endothelial origin, SK-HEP-1 remains widely employed in hepatocellular carcinoma and liver cancer research, particularly for investigating tumor cell migration, invasion, and drug responsiveness.
CCDC85C functions as a p53-inducible tumor suppressor that directly links p53 activation to Hippo pathway regulation. Upon genotoxic stress, p53 transcriptionally upregulates CCDC85C, which physically interacts with the transcriptional coactivator YAP. This interaction recruits the FBXW7 E3 ubiquitin ligase, leading to YAP ubiquitination and proteasomal degradation. Reduced YAP abundance limits the formation of YAP/TEAD transcriptional complexes, thereby suppressing the expression of downstream targets such as CTGF and CYR61. Concurrently, CCDC85C promotes apoptosis through the induction of proapoptotic factors like BAX and PUMA. Thus, CCDC85C serves as a critical effector of p53-mediated growth inhibition by bridging DNA damage responses to YAP-dependent proliferative and survival cues.
In the SK-HEP-1 metastatic adenocarcinoma model, knockout of CCDC85C allows researchers to examine the consequences of impaired p53-YAP cross talk on cellular behaviors relevant to liver cancer progression. The loss of this tumor suppressor is expected to enhance YAP transcriptional activity, potentially accelerating cell proliferation, migration, and resistance to apoptosis. This model is particularly suited for exploring how disruption of the CCDC85C-YAP axis contributes to the aggressive phenotype of liver adenocarcinoma and for identifying context-dependent dependencies that may inform therapeutic strategies. The cell line??s background also offers a platform to evaluate the interplay between CCDC85C and other signaling pathways commonly dysregulated in metastatic disease.
Typical applications include western blotting to assess CCDC85C, p53, and YAP protein levels; RT-qPCR for monitoring CTGF and CYR61 expression; co-immunoprecipitation to validate CCDC85C?CYAP interactions; and functional assays such as Annexin V/PI staining for apoptosis, cell viability measurements, and migration or invasion assays. Immunofluorescence can be used to observe YAP subcellular localization changes. These assays support investigations into p53-mediated tumor suppression, Hippo pathway regulation, and liver cancer biology, as well as drug screening efforts targeting YAP transcriptional activity. For additional technical details and support, please contact Ascent Research.
