The GSK3B Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the SK-HEP-1 human hepatocellular carcinoma line, engineered for disruption of the glycogen synthase kinase 3 beta (GSK3B) gene. This polyclonal knockout pool provides a heterogeneous cell population lacking functional GSK3B, enabling robust investigation of GSK3B-dependent signaling networks in a liver cancer context. The knockout model is generated without single-cell cloning, making it suitable for experiments requiring population-level loss-of-function analysis.
SK-HEP-1 is a well-characterized hepatocellular carcinoma cell line originally isolated from the ascitic fluid of a 52-year-old male patient with liver adenocarcinoma. It is extensively used to model hepatic tumorigenesis, metabolic reprogramming, and drug evaluation in liver cancer research. Its hepatocellular phenotype and retained metabolic functions make it particularly valuable for studying kinase-mediated regulation of glycogen metabolism and oncogenic signaling. The GSK3B knockout in this background allows precise interrogation of tumor cell biology within a clinically relevant genetic and metabolic framework.
GSK3B is a serine/threonine kinase that functions as a central regulator of multiple signaling cascades, including Wnt/??-catenin, PI3K/Akt, and insulin pathways. Under basal conditions, GSK3B associates with AXIN1, APC, and ??-catenin in a destruction complex, phosphorylating ??-catenin to promote its ubiquitin-mediated proteolysis. Knockout of GSK3B eliminates this negative regulation, resulting in ??-catenin stabilization, nuclear accumulation, and TCF/LEF-driven transcription of targets such as c-Myc and cyclin D1. Upstream kinases AKT and PKA inhibit GSK3B through phosphorylation, while Wnt ligands, Frizzled receptors, and LRP6/DVL2 signaling oppose its activity. GSK3B also interacts with FRAT1 and PP2A and phosphorylates glycogen synthase, linking metabolic and proliferative pathways.
In hepatocellular carcinoma, aberrant Wnt/??-catenin signaling is a frequent oncogenic driver, and GSK3B knockout in SK-HEP-1 cells mimics this pathological activation. The resulting constitutive ??-catenin signaling promotes uncontrolled proliferation, resistance to apoptosis, and transcriptional programs associated with metastasis. The polyclonal nature preserves cellular heterogeneity, which may better reflect tumor diversity and response to perturbations, thereby facilitating studies of ??-catenin-mediated oncogenesis and identification of context-specific vulnerabilities.
Representative applications include quantitative immunofluorescence for ??-catenin nuclear translocation, ??-catenin luciferase reporter assays to measure TCF/LEF activity, and RT-qPCR profiling of key target genes. The cells are amenable to high-throughput drug screening for GSK3?? inhibitors, as well as proliferation (MTT) and apoptosis assays. Phospho-specific western blotting enables concurrent evaluation of AKT/PKA signaling. This polyclonal knockout product provides a versatile platform for hepatocellular carcinoma research and Wnt pathway dissection. For detailed product information or customized services, please contact Ascent Research.
