The CBX8 Knockout Huh-7 Polyclonal Cells provided by Ascent Research are a CRISPR/Cas9-edited polyclonal cell population in which the CBX8 gene has been disrupted through targeted gene editing, offering a robust loss-of-function model for investigating the role of CBX8 in hepatocellular carcinoma and epigenetic regulation. The polyclonal nature of this knockout pool captures the diverse editing outcomes within the cell population and is well-suited for functional assays without the need for single-cell-derived clones. This model enables researchers to dissect the molecular consequences of CBX8 deficiency in a medically relevant liver cancer background.
Derived from a well-differentiated hepatocellular carcinoma, the Huh-7 cell line is a widely used model of liver cancer, originally established from the tumor of a male patient. These adherent epithelial cells retain key characteristics of hepatocytes and are amenable to high-efficiency genome editing and downstream biochemical assays. Their robust growth and responsiveness to Wnt/??-catenin signaling make them an ideal host for interrogating the function of chromatin-modifying factors such as CBX8, whose aberrant expression is implicated in hepatocarcinogenesis.
CBX8 encodes a chromodomain-containing subunit of Polycomb repressive complex 1 (PRC1) that specifically recognizes trimethylated lysine 27 on histone H3 (H3K27me3), a repressive chromatin mark deposited by EZH2. Through its interaction with RING1B and BMI1, CBX8 facilitates the monoubiquitination of histone H2A at lysine 119 (H2AK119ub), promoting chromatin compaction and transcriptional silencing. In hepatocellular carcinoma, CBX8 overexpression is transcriptionally activated by the ??-catenin/TCF complex and E2F1, and it further represses key tumor suppressors such as CDKN2A and CDKN1A, as well as Wnt pathway inhibitors including DKK1 and SFRP1. This feedforward mechanism potentiates Wnt/??-catenin signaling, driving expression of pro-proliferative and stemness-associated genes downstream of TCF4. CBX8 also physically associates with PHC proteins and is regulated by miR-185-3p, integrating multiple oncogenic inputs.
Within Huh-7 cells, CBX8 knockout disrupts this oncogenic axis, making the polyclonal knockout population a powerful tool for studying epigenetic dependencies in liver cancer. Loss of CBX8 is expected to derepress tumor suppressor loci and dampen Wnt/??-catenin transcriptional output, providing a platform to evaluate its role in proliferation, apoptosis resistance, and maintenance of the cancer stem cell phenotype. The model also allows investigation of interactions with other PRC1 components and its influence on H3K27me3-associated gene regulation in a hepatic context.
Researchers can employ these CBX8 knockout cells in a wide range of experimental applications, including RNA-seq and ChIP-qPCR to map changes in the epigenetic landscape and transcriptome, Western blotting and immunofluorescence to confirm protein-level alterations, and flow cytometry or migration assays to assess phenotypic shifts. The cells are also suitable for drug sensitivity screens to identify synthetic lethal vulnerabilities or to test small molecules targeting the Wnt/??-catenin pathway, using luciferase-based TCF/LEF reporters. For detailed protocols or to place an order, please contact Ascent Research.
