The KCTD12 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population targeting the KCTD12 gene in the human SK-HEP-1 liver adenocarcinoma cell line. This product provides a loss-of-function model to study KCTD12-dependent signaling and ubiquitination mechanisms. Generated through CRISPR/Cas9-mediated gene disruption, the polyclonal pool enables robust analysis of gene function without clone-specific artifacts, offering a versatile tool for functional studies in hepatocellular carcinoma research. The knockout cells are suitable for a variety of downstream applications that require ablated KCTD12 expression in a hepatic cancer background.
The host SK-HEP-1 cell line, originally derived from the ascites of a patient with liver adenocarcinoma, is a widely utilized model for hepatocellular carcinoma (HCC) research, metastasis studies, and drug screening. SK-HEP-1 cells exhibit an epithelial morphology and retain key features of liver cancer cells, including aggressive growth and metastatic potential. This cell line has been extensively employed to investigate the molecular mechanisms underlying HCC progression, chemoresistance, and tumor microenvironment interactions. The genetic knockout of KCTD12 in this established background permits dissection of its specific contributions to liver cancer cell behavior.
KCTD12 functions as a substrate adaptor for the Cullin3 (CUL3)-RING E3 ubiquitin ligase complex, directing the ubiquitination of GABAB receptor subunits GABBR1 and GABBR2 and controlling their subsequent trafficking and signaling. This interaction is crucial for modulating GABAB receptor-mediated inhibition of adenylyl cyclase, which suppresses cAMP production and downregulates protein kinase A (PKA) activity. KCTD12 also associates with other adaptors such as KCTD16 and is regulated by cAMP and PKA, forming a feedback loop that fine-tunes receptor surface availability and downstream effectors like KCNJ potassium channels. Disruption of KCTD12 therefore alters the balance of GABAB receptor signaling, impacting cAMP/PKA pathway dynamics and cellular excitability.
In the context of hepatocellular carcinoma, aberrant GABAB receptor signaling and cAMP/PKA pathway activity have been implicated in tumor growth, invasion, and drug resistance. By creating a KCTD12 knockout in SK-HEP-1 cells, researchers can investigate how loss of this adaptor affects ubiquitination-dependent receptor trafficking and downstream signaling cascades that drive liver cancer cell proliferation and metastasis. This model is particularly valuable for dissecting the role of KCTD12 in modulating cellular responses to extracellular stimuli and chemotherapeutic agents, providing insights into potential therapeutic targets within the ubiquitin-proteasome system.
The KCTD12 Knockout SK-HEP-1 Polyclonal Cells are suitable for a wide range of research applications, including liver cancer biology, GABAB receptor signaling, ubiquitination mechanisms, and drug resistance studies. Representative assays supported by this model include Western blotting for protein expression analysis, RT-qPCR for transcriptional profiling, immunofluorescence for subcellular localization, cAMP assays to measure second messenger levels, cell viability and migration/invasion assays for functional phenotyping, and co-immunoprecipitation to study protein-protein interactions. For additional technical details or custom orders, please contact Ascent Research.
