The HS1BP3 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the human hepatic adenocarcinoma cell line SK-HEP-1, designed for targeted disruption of the HS1BP3 gene. This product consists of a heterogeneous pool of edited cells, enabling comprehensive loss-of-function studies while maintaining the genetic diversity inherent to tumor cell populations. By circumventing monoclonal selection, these polyclonal knockout cells better represent the complexity of cancer biology, facilitating robust functional assays in a liver cancer context.
SK-HEP-1 is a widely employed cell line originally established from the ascites of a patient with liver adenocarcinoma. Although classified as adenocarcinoma, SK-HEP-1 exhibits endothelial-like protein expression and mesenchymal features, and is broadly utilized as a hepatocellular carcinoma model. Its aggressive phenotype and rapid growth make it a versatile platform for studying tumor biology, metastasis, and drug responses in hepatic malignancies.
HS1BP3 encodes an adaptor protein central to clathrin-mediated endocytosis, interacting with clathrin heavy chain, AP-2 complex subunits, dynamin, and HCLS1 to regulate receptor internalization and trafficking. It facilitates EGFR endocytosis and endosomal sorting, balancing receptor recycling and degradation. Knockout of HS1BP3 disrupts ligand-induced EGFR internalization, attenuating downstream signaling. Mechanistically, HS1BP3 operates downstream of EGF, TGF-alpha, and Src kinases, and its loss impairs AKT and ERK1/2 activation, reducing proliferation and migration. HS1BP3 is thus a critical mediator of EGFR signaling.
In the SK-HEP-1 background, disruption of HS1BP3 perturbs EGFR-driven oncogenic signaling, frequently dysregulated in liver cancer. The mesenchymal and metastatic characteristics of SK-HEP-1 make this knockout model valuable for dissecting the contribution of endocytic trafficking to tumor aggressiveness and therapy resistance. Altered EGFR turnover may sensitize cells to erlotinib, offering a platform for studying combination therapies that target endocytic vulnerabilities in hepatic adenocarcinoma.
Researchers can use these cells to study EGFR internalization via fluorescent ligand uptake, and analyze signaling by Western blot for phospho-EGFR, p-ERK, and p-AKT. Immunofluorescence can assess clathrin-EGFR co-localization. Functional assays include MTT proliferation, wound healing migration, and Annexin V apoptosis. The model is suitable for drug sensitivity screening with EGFR inhibitors and RNA-seq transcriptomic profiling to uncover global changes upon HS1BP3 loss. It also serves as a system for investigating clathrin-mediated endocytosis in hepatic adenocarcinoma. For further information, please contact Ascent Research.
