The AXL Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population featuring disruption of the AXL gene. This heterogeneous pool offers a robust loss-of-function model that avoids clonal selection artifacts and captures population-level effects, ensuring reliable ablation of AXL-dependent signaling. The polyclonal format provides physiologically relevant conditions for studying the receptor??s role in liver cancer cell biology.
The host cell line, SK-HEP-1, was established from the ascites of a patient with hepatic adenocarcinoma and is a widely employed model for hepatocellular carcinoma (HCC). These cells display endothelial-like characteristics, including the ability to form capillary-like structures, making them particularly suited for investigations into tumor angiogenesis, metastasis, and the epithelial-to-mesenchymal transition (EMT). Introduction of the AXL knockout into this background creates a powerful platform for dissecting oncogenic signaling.
AXL encodes a receptor tyrosine kinase that transduces extracellular signals from the ligands Gas6 and Protein S. Ligand binding induces AXL homodimerization and autophosphorylation, creating docking sites for adaptors such as Grb2 and the p85 regulatory subunit of PI3K. This activates downstream cascades including PI3K/AKT, MAPK/ERK, JAK/STAT, and NF-??B. Key transcriptional targets that promote survival and invasion are Survivin, Bcl-2, Cyclin D1, MMP-9, Snail, and Twist. AXL expression is induced by hypoxia, TNF-??, and IL-6, and its signaling is tempered by SOCS proteins. Disruption of AXL prevents Gas6-stimulated phosphorylation of AKT and ERK1/2, blocks NF-??B nuclear translocation, and thereby silences the gene programs driving proliferation and migration.
In SK-HEP-1 cells, functional loss of AXL dramatically reduces Gas6-mediated cell viability and motility, underscoring the receptor??s importance in HCC progression. The endothelial-like phenotype also enables studies of AXL in vascular mimicry and tumor-endothelial crosstalk. This polyclonal knockout population is an essential resource for validating anti-AXL therapies, as it eliminates confounding residual receptor activity and models the heterogeneity observed in clinical tumors.
Key applications include signal transduction analysis, liver cancer metastasis research, drug resistance mechanism studies, and preclinical target validation. Researchers can utilize these cells in Western blotting for total AXL and phospho-AKT, RT-qPCR for AXL mRNA, transwell migration and invasion assays, flow cytometric apoptosis and cell cycle analysis, MTT or CellTiter-Glo viability assays, and immunofluorescence staining for EMT markers like vimentin and Snail. This product supports rigorous interrogation of AXL biology in a clinically relevant hepatic adenocarcinoma model. For additional information, please contact Ascent Research.
