The ITSN1 Knockout SK-HEP-1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ITSN1 gene in the human SK-HEP-1 hepatic adenocarcinoma cell line. This polyclonal format comprises a heterogeneous pool of cells with diverse gene-editing events, providing a robust loss-of-function model that minimizes clonal bias and ensures reproducible functional studies. CRISPR/Cas9-mediated gene disruption stably impairs ITSN1 expression, allowing researchers to dissect the scaffold protein??s roles in a defined genetic background.
SK-HEP-1 cells were originally isolated from the ascitic fluid of a 52-year-old male with liver adenocarcinoma. They uniquely co-express epithelial and endothelial markers, making them a valuable model for studying hepatocellular carcinoma metastasis, tumor cell plasticity, and vasculogenic mimicry. This cell line is widely employed to investigate mechanisms of migration, invasion, and the interaction of cancer cells with their microenvironment.
ITSN1 encodes a multidomain scaffold protein that integrates endocytosis, receptor trafficking, and actin cytoskeleton dynamics. Activated downstream of EGFR, Src family kinases, and PI3K/AKT signaling, ITSN1 recruits epsin, dynamin, synaptojanin, and PI3K-C2?? to facilitate clathrin-mediated internalization, while its SH3 domains bind SOS1 to activate the Ras?CRaf?CMEK?CERK cascade. Additionally, ITSN1 engages Cbl to modulate receptor ubiquitination and endosomal sorting, and promotes actin polymerization via Cdc42 and N-WASP. This coupling of EGFR internalization to MAPK/ERK signaling drives cell proliferation and migration, positioning ITSN1 as a key nexus in oncogenic signal transduction.
In the SK-HEP-1 hepatic adenocarcinoma model, ITSN1 knockout enables dissection of how scaffold loss alters EGFR trafficking, MAPK/ERK output, and cytoskeletal remodeling in a metastatic liver cancer context. The dual epithelial?Cendothelial nature of these cells is particularly advantageous for examining ITSN1??s contributions to both tumor cell?Cautonomous functions and endothelial-like behaviors that facilitate dissemination. This system can reveal context-specific roles of ITSN1 in hepatocellular carcinoma progression and adaptive signaling rewiring following gene disruption.
This polyclonal knockout cell population supports a range of experimental applications, including immunofluorescence-based EGFR localization and trafficking assays, EGF-stimulated phospho-ERK quantification, endocytosis uptake measurements, and cell migration/invasion assays to assess metastatic potential. Cell proliferation assays further support investigations into growth dependency, while drug discovery efforts can employ this model for screening ITSN1-targeted inhibitors. For additional technical details and ordering information, please contact Ascent Research.
