The BROX Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the SK-HEP-1 human liver adenocarcinoma cell line, engineered to disrupt BROX gene function. This heterogeneous pool of edited cells ensures diverse loss-of-function alleles without clonal selection, providing a robust model for studying dynamic processes such as abscission and autophagy. The polyclonal format minimizes clone-specific artifacts and enables population-based assays with enhanced statistical relevance.
SK-HEP-1 cells, isolated from ascitic fluid of a male patient with liver adenocarcinoma, exhibit both epithelial and endothelial properties, making them a versatile model for hepatocellular carcinoma, tumor angiogenesis, and vascular mimicry. Their malignant characteristics and maintenance of liver cancer-relevant signaling pathways provide a biologically appropriate context for examining BROX-dependent membrane trafficking and cell division.
BROX encodes a membrane-anchored adaptor with an N-terminal BRO1 domain and a C-terminal CAAX prenylation motif that directs localization to the midbody. It functions downstream of cell cycle cues to recruit CHMP4B, nucleating ESCRT-III filament assembly together with CHMP2A and VPS4. Interaction with ALIX (PDCD6IP) further stabilizes this complex at sites of membrane fission. BROX thus orchestrates membrane scission during abscission and is also required for autophagosome closure, linking ESCRT-III function to both mitosis and degradative autophagy.
Loss of BROX in SK-HEP-1 cells causes abscission failure, yielding multinucleated phenotypes that serve as quantifiable readouts for cytokinetic defects and their contribution to chromosomal instability in liver cancer. Impaired autophagic flux in these cells can influence metabolic adaptation and drug sensitivity, making the model relevant for studying chemoresistance mechanisms. The polyclonal nature ensures broad representation of phenotypes, enhancing reproducibility in mechanistic studies.
These polyclonal cells are applicable in multinucleation and abscission assays monitored by live-cell imaging or immunofluorescence. Co-immunoprecipitation and western blotting using these cells enable analysis of CHMP4B, ALIX, and VPS4 recruitment and ESCRT-III complex integrity. Autophagy flux assays with LC3 reporters further reveal roles of BROX in membrane trafficking. This knockout tool supports high-content screening and detailed dissection of ESCRT-mediated pathways in cancer cell biology. For technical inquiries, contact Ascent Research.
