The CCDC88B knockout SK-HEP-1 polyclonal cells constitute a CRISPR/Cas9-edited human cell population in which the CCDC88B gene has been disrupted. This polyclonal knockout model is generated from the SK-HEP-1 host cell line and provides a genetically heterogenous pool of cells carrying diverse loss-of-function mutations in the target locus. Such a format avoids the clonal artifacts associated with single-cell?Cderived knockouts and is particularly useful for studying gene function in a population-level context, reflecting the inherent variability of tumor biology. The product is designed for researchers investigating non-canonical Wnt signaling, cell migration, and planar cell polarity mechanisms in a hepatic adenocarcinoma background.
SK-HEP-1 is a human cell line originally isolated from the ascites of a patient with liver adenocarcinoma. Although widely employed as a surrogate for liver sinusoidal endothelial cells, it retains adenocarcinoma characteristics and expresses both epithelial and mesenchymal markers. The cells exhibit an adherent, endothelial-like morphology and are frequently used in studies of hepatocellular carcinoma biology, metastasis, and drug response. Their dual epithelial?Cmesenchymal phenotype makes them an ideal host for examining genes involved in cell polarity, migration, and cytoskeletal dynamics, as these processes are critical in cancer progression and tissue morphogenesis.
CCDC88B encodes a scaffold protein that serves as an obligate adaptor in the non-canonical Wnt/planar cell polarity pathway. Mechanistically, it directly interacts with Dishevelled proteins (DVL1, DVL2, DVL3) downstream of Frizzled receptor activation by ligands such as Wnt5a and Wnt11. This facilitates the localized activation of the small GTPase RhoA, leading to Rho-associated kinase (ROCK)?Cdependent actin remodeling and JNK-mediated transcriptional responses via AP-1. CCDC88B also associates with GRP78 (HSP5A) and participates in ciliogenesis. Thus, it functions as a molecular hub linking extracellular polarity cues to cytoskeletal reorganization and gene expression, ultimately controlling directional cell movement and tissue patterning.
Disruption of CCDC88B in the SK-HEP-1 background creates a relevant model for dissecting the molecular requirements of non-canonical Wnt signaling in a metastatic adenocarcinoma context. Loss of CCDC88B is predicted to impair RhoA activation and JNK phosphorylation, leading to reduced actin polymerization and defective cell migration and invasion. This directly mimics aspects of tumor cell dissemination and inflammatory cell trafficking, where planar cell polarity signaling is often dysregulated. The model is especially pertinent for studying CCDC88B-related pathologies such as Crohn??s disease and ciliopathies, as well as for exploring therapeutic strategies aimed at modulating Wnt/RhoA/JNK axis.
Typical experimental applications include wound healing and Transwell migration/invasion assays to quantify directional motility defects. RhoA activation can be measured directly via G-LISA, while phospho-JNK levels are assessed by western blotting. Phalloidin staining reveals actin cytoskeleton perturbations, and AP-1 luciferase reporters monitor downstream transcriptional activity. Co-immunoprecipitation experiments with DVL isoforms confirm disrupted protein?Cprotein interactions. Flow cytometry may be used to evaluate changes in cell surface marker expression. For further technical details or custom project inquiries, please contact Ascent Research.
