The DLG1 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 comprises a heterogeneous pool of cells harboring diverse disruption events at the DLG1 locus, generated by CRISPR/Cas9-mediated gene targeting. The polyclonal format provides a robust loss-of-function model without the clonal selection artifacts often associated with single-cell-derived lines. The knockout population is validated at the protein level and is suitable for functional studies of DLG1 in hepatocellular carcinoma and related epithelial cancers.
The SK-HEP-1 host cell line originated from the ascites of a patient with liver adenocarcinoma. These cells exhibit an unusual co-expression of epithelial and endothelial markers, reflecting a dedifferentiated phenotype typical of aggressive hepatocellular carcinoma. SK-HEP-1 cells grow as an adherent monolayer and have been extensively used to investigate hepatic tumor biology, metastasis, and drug responses. Their rapid proliferation and ease of manipulation make them a valuable platform for genetic perturbation studies, particularly for genes involved in cell polarity and signaling.
DLG1 (Discs Large Homolog 1) is a member of the membrane-associated guanylate kinase (MAGUK) family and functions as a multidomain scaffolding protein essential for maintaining apical-basal polarity and organizing intercellular junctions. DLG1 forms complexes with the aPKC-PAR3-PAR6 and Scribble-Lgl polarity modules, and interacts directly with tight junction components such as CLDN1 and OCLN. In the Wnt pathway, DLG1 associates with APC and ??-catenin, regulating ??-catenin stability and transcriptional activity at TCF/LEF target genes like MYC and CCND1. In the Hippo pathway, DLG1 partners with Scribble to promote MST1/2 and LATS activation, restraining YAP/TAZ activity and reducing expression of pro-proliferative genes such as CTGF and CYR61. The scaffold also interfaces with PTEN and KRAS, integrating signals that govern epithelial homeostasis.
In SK-HEP-1 hepatocellular carcinoma cells, DLG1 knockout disrupts the coordination between cell polarity and growth signaling, recapitulating molecular events frequently observed in liver cancer progression. Loss of DLG1 destabilizes tight junctions and compromises contact inhibition, leading to constitutive activation of both ??-catenin and YAP/TAZ transcriptional programs. This results in enhanced proliferation, migration, and invasion??hallmarks of aggressive malignancy. The polyclonal knockout population provides a physiologically relevant model to study how loss of a single polarity scaffold can concurrently activate multiple oncogenic pathways, mirroring the multigenic nature of hepatocellular carcinoma.
This knockout model is suitable for mechanistic studies of DLG1 signaling crosstalk, drug screening for polarity-restoring compounds, and functional genomics of MAGUK scaffolds. Assays include Western blotting for DLG1 and downstream targets, immunofluorescence for tight junction markers (CLDN1, ZO-1), TOPFlash/FOPFlash and YAP/TAZ luciferase reporters, Transwell migration and invasion tests, and co-immunoprecipitation of DLG1-??-catenin complexes. RNA-seq enables transcriptome-wide profiling. For more details, contact Ascent Research.
