The CCM2 Knockout SK-HEP-1 Polyclonal Cells comprise a CRISPR/Cas9-mediated gene-disrupted population derived from the SK-HEP-1 human liver adenocarcinoma cell line. This product delivers a heterogeneous knockout pool that preserves genetic diversity, providing a versatile platform for studying CCM2 loss-of-function effects. The polyclonal format eliminates the need for single-cell cloning while supporting high-throughput and pooled-assay applications. These cells enable investigation of CCM2 signaling in a model that bridges hepatic cancer biology and endothelial vascular research.
SK-HEP-1 cells were originally isolated from a liver adenocarcinoma patient and exhibit adherent epithelial morphology. Despite their hepatic origin, they are frequently employed as a surrogate endothelial model due to expression of endothelial markers and responsiveness to angiogenic stimuli. This dual identity makes them especially useful for dissecting molecular pathways governing vascular junction integrity and tumor?Cmicroenvironment interactions. In the CCM2 knockout context, SK-HEP-1 cells offer a pertinent system to examine mechanisms of endothelial barrier regulation and cytoskeletal dynamics.
CCM2 encodes a scaffold protein that forms a ternary complex with KRIT1 (CCM1) and PDCD10 (CCM3), orchestrating endothelial junction stability and actin dynamics. This complex is regulated by integrin beta1 and VEGF receptor signaling, and it directly suppresses RhoA GTPase activity to limit ROCK1/2-mediated cytoskeletal tension. CCM2 also engages MEKK3 kinase to modulate MAPK pathway output, while interacting with STK25 and focal adhesion kinase (FAK) at adhesion sites. Loss of CCM2 ablates these protein?Cprotein interactions, resulting in constitutive RhoA-ROCK activation, disassembly of adherens junctions, and aberrant transcriptional activation driven by MEKK3-dependent signaling.
In SK-HEP-1 cells, CCM2 knockout disrupts epithelial integrity and promotes a hypercontractile phenotype, mimicking endothelial dysfunction seen in cerebral cavernous malformations. This polyclonal model enables investigation of RhoA-dependent permeability defects and the reconstitution of CCM complex interactions. Because SK-HEP-1 cells retain endothelial-like plasticity, they serve as a practical platform for probing vascular stability mechanisms and testing rescue by ROCK inhibitors. The model thus bridges oncological and vascular research, facilitating dissection of CCM2??s tumor-suppressive and barrier-protective roles.
Key applications include Western blotting for junctional proteins (VE-cadherin, ZO-1), RhoA-GTP pull-down assays, immunofluorescence analysis of actin stress fibers, and endothelial permeability assays. The polyclonal format is compatible with pooled RNA-seq, co-immunoprecipitation of CCM complex members, and high-throughput screening of ROCK pathway inhibitors. Drug sensitivity studies using this model can identify compounds that restore vascular stability in CCM-related disorders. For further technical details or experimental consultation, please contact Ascent Research.
