The IRS1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the insulin receptor substrate 1 (IRS1) gene in a human hepatic adenocarcinoma background. This product consists of a mixed population of edited cells with targeted disruption of the IRS1 locus, generated by CRISPR/Cas9-mediated genome editing, providing a robust experimental system for investigating IRS1-dependent signaling without the variability of single-cell clones.
The SK-HEP-1 cell line is a widely employed model derived from the ascitic fluid of a patient with liver adenocarcinoma. These cells exhibit endothelial-like properties alongside their hepatic tumor origin, making them a valuable tool for studying liver cancer biology, metastasis, and the interplay between metabolic and oncogenic signaling pathways. SK-HEP-1 cells retain key features of hepatocyte and vascular endothelial cells, enabling multifaceted experimental designs.
IRS1 functions as a crucial cytoplasmic adaptor protein in insulin and insulin-like growth factor 1 (IGF-1) signaling. Upon ligand binding, tyrosine phosphorylation of IRS1 by upstream receptor tyrosine kinases such as INSR and IGF1R creates docking sites for SH2 domain-containing proteins, notably the p85 regulatory subunit of PI3K and the adaptor GRB2. This event triggers activation of the PI3K-AKT cascade, involving intermediaries like PDK1 and mTORC2, and the MAPK/ERK pathway via SHC-GRB2-SOS. Downstream effectors include AKT, mTORC1, S6K, GSK3??, and FOXO transcription factors, which collectively regulate glucose metabolism, protein synthesis, cell proliferation, and survival. Additionally, IRS1 interacts with PTP1B and JAK2, and its stability and function are modulated by upstream regulators such as TNF-??, IL-6, JNK, and IKK-??, linking inflammatory and stress signals to metabolic control.
In the context of SK-HEP-1 cells, IRS1 knockout provides a powerful model to dissect the interplay between insulin signaling and hepatic tumor biology. Given that SK-HEP-1 cells exhibit both endothelial and adenocarcinoma characteristics, loss of IRS1 can help elucidate how metabolic reprogramming driven by PI3K-AKT-mTOR and MAPK pathways contributes to hepatocellular carcinoma progression, angiogenesis, and therapy resistance. This model is particularly relevant for studying the dual roles of insulin signaling in liver metabolism and tumorigenesis, including the investigation of insulin resistance, metabolic syndrome, and obesity-associated liver cancer.
Researchers can employ the IRS1 Knockout SK-HEP-1 Polyclonal Cells in a diverse array of functional assays. Typical applications include Western blotting and phospho-signaling analyses to assess changes in AKT, ERK1/2, and GSK3?? phosphorylation, glucose uptake assays to quantify metabolic alterations, and proliferation or apoptosis assays to evaluate cell fate decisions. This model is also suitable for RNA-seq and metabolic flux analyses to profile global transcriptomic and metabolic shifts upon IRS1 disruption. Moreover, these cells can serve as a critical tool for screening small molecules or biologics that modulate IRS1 function or bypass IRS1-dependent signaling, benefiting drug discovery efforts in diabetes, obesity, and liver cancer. For further information or technical support, please contact Ascent Research.
