The ASAP2 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human liver adenocarcinoma cell line. This product features targeted disruption of the ASAP2 gene (DDEF2) across a heterogeneous pool, providing a versatile loss-of-function model for studying this Arf6-specific GTPase-activating protein in a hepatocellular carcinoma context.
The parental SK-HEP-1 cell line is an adherent, epithelial-like line isolated from ascites of a liver adenocarcinoma patient and displays an aneuploid karyotype. It is widely used as a model for liver cancer and hepatocyte biology, retaining malignant features including migratory and invasive potential. Integrating the ASAP2 knockout into this background creates a physiologically relevant platform for dissecting pathways that drive tumor cell motility and metastasis.
ASAP2 inactivates the small GTPase Arf6 by accelerating GTP hydrolysis, thereby attenuating Arf6-mediated actin cytoskeleton reorganization, lamellipodia formation, and cell migration. Upstream, EGF and receptor tyrosine kinases stimulate PI3K, which can regulate ASAP2. Downstream, ASAP2-mediated Arf6 inactivation modulates Rac1 activity, actin remodeling, and focal adhesion disassembly. ASAP2 interacts with cortactin and paxillin, linking it to actin dynamics. Key pathway components include Arf6, ASAP2, Rac1, PAK1, and cortactin. Disruption of ASAP2 is expected to cause sustained Arf6 activation and enhanced invasive behavior.
In SK-HEP-1 cells, loss of ASAP2 likely perturbs Arf6-dependent cytoskeletal regulation, augmenting migration and invasion. Given that aberrant Arf6 signaling is linked to metastasis, this knockout model enables investigation of ASAP2??s role in hepatocellular carcinoma progression. The aneuploid background may interact with ASAP2 loss, offering a model for studying combinatorial effects on cancer cell behavior and genomic instability.
The cells are suited for functional assays such as wound healing, transwell invasion, and 3D spheroid invasion to quantify motility. Biochemical validation includes Western blotting, GTPase activity assays, and co-immunoprecipitation of ASAP2 partners like cortactin and paxillin. Immunofluorescence can visualize actin and focal adhesions, while qPCR and screening approaches identify pathway modulators. For further details, contact Ascent Research.
