ACTN1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from human SK-HEP-1 liver adenocarcinoma cells, featuring disruption of the ACTN1 gene. The heterogeneous polyclonal pool provides a robust loss-of-function model that avoids clonal artifacts, allowing reliable interrogation of ACTN1 roles in cell adhesion, migration, and mechanotransduction.
SK-HEP-1 is a mesenchymal-like human liver adenocarcinoma cell line with a spindle morphology and high invasive capacity, originally isolated from ascitic fluid. These cells co-express endothelial and epithelial markers, making them a valuable model for investigating endothelial-mesenchymal transition and hepatocellular carcinoma metastasis. The SK-HEP-1 background offers a clinically relevant context for studying ACTN1’s contributions to tumorigenic and metastatic processes.
Alpha-actinin-1 (ACTN1) functions as a homodimeric actin-bundling protein that crosslinks F-actin filaments and anchors them to focal adhesions through direct binding to integrin beta1, vinculin, talin, zyxin, paxillin, and PIP2. Its activity is regulated upstream by integrin-mediated adhesion, FAK, Src family kinases, TGF-??, Rho GTPases (RhoA, Rac1, Cdc42), and mechanical tension. Downstream, ACTN1-organized actin networks drive focal adhesion turnover, SRF transcriptional activity, and YAP/TAZ nuclear translocation, thereby coupling cytoskeletal mechanics to pro-migratory and pro-proliferative gene expression. In hepatocellular carcinoma, ACTN1 integrates signals from the extracellular matrix to promote metastasis via MAPK/ERK signaling, enhancing invasive capacity.
Disruption of ACTN1 in these SK-HEP-1 polyclonal knockout cells compromises the structural coupling between the actin cytoskeleton and integrin-based adhesions, resulting in impaired cell spreading, reduced migration, and diminished invasiveness. Given the mesenchymal nature of SK-HEP-1, the model is well-suited for dissecting amoeboid-to-mesenchymal transition and collective invasion. Overexpression of ACTN1 is associated with poor prognosis in hepatocellular carcinoma and colorectal cancer, making this knockout a relevant tool for studying metastatic vulnerabilities. Loss of ACTN1 is expected to attenuate mechanosensitive YAP/TAZ signaling, thereby suppressing target genes such as CTGF and CYR61, which are critical for tumor-stroma crosstalk and angiogenesis.
This polyclonal knockout product is compatible with a wide array of assays. Western blotting and immunofluorescence microscopy enable verification of ACTN1 depletion and assessment of focal adhesion proteins like vinculin and paxillin. Transwell migration/invasion, wound healing, and cell adhesion assays quantify phenotypic changes, while phospho-signaling analysis of FAK and YAP reveals mechanotransduction alterations. Live-cell imaging of fluorescently tagged adhesions can capture dynamic adhesion turnover. The model also supports anti-metastatic drug screening and siRNA complementation studies. For further information, please contact Ascent Research.
