The GRB10 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population designed for the loss-of-function study of GRB10 in a human liver adenocarcinoma background. Generated using CRISPR/Cas9 technology, this polyclonal knockout model introduces targeted disruptions in the GRB10 gene within the SK-HEP-1 host cell line, resulting in a heterogeneous pool of cells carrying diverse GRB10-inactivating alleles. This product format enables robust phenotypic screening and downstream molecular analyses without the clonal selection artifacts associated with single-cell-derived lines.
The SK-HEP-1 cell line was originally derived from the ascitic fluid of a patient with liver adenocarcinoma and is notable for its atypical endothelial-like characteristics, including cobblestone morphology and expression of endothelial markers such as von Willebrand factor. This dual identity??possessing both epithelial tumorigenic properties and endothelial features??makes SK-HEP-1 a unique platform for investigating hepatocellular carcinoma progression and the molecular underpinnings of tumor angiogenesis. The line??s well-characterized signaling networks and adaptable growth conditions further support its utility in cancer biology research.
GRB10 is an adaptor protein that negatively regulates insulin and insulin-like growth factor 1 (IGF-1) signaling. Upon ligand stimulation, GRB10 binds to phosphorylated tyrosine residues on activated INSR and IGF1R, blocking recruitment of IRS1 and IRS2. This inhibits downstream PI3K/AKT and MAPK/ERK cascades, thereby attenuating cell proliferation, survival, and metabolic responses. Its activity is modulated by insulin/IGF-1, FOXO transcription factors, and stress signals, while interactions with NEDD4, 14-3-3 proteins, and GIGYF2 contribute to receptor-proximal signaling dynamics.
In the SK-HEP-1 background, GRB10 disruption relieves tonic inhibition of insulin/IGF-1 pathways, leading to hyperactivation of PI3K/AKT/mTOR and MAPK/ERK signaling. Given the line??s hepatic and endothelial-like features, this model dissects how enhanced growth factor signaling drives liver cancer cell proliferation, migration, and angiogenic potential. Researchers can also investigate GRB10??s role in vascular biology, including endothelial cell survival and tube formation, within a cancer-relevant setting. This knockout thus enables study of oncogenic amplification effects on tumor-autonomous and microenvironmental processes.
These cells are suitable for insulin/IGF-1 stimulation assays, Western blotting for phospho-AKT (Ser473), phospho-ERK (Thr202/Tyr204), and GRB10, and RT-qPCR of target genes. Functional studies include cell proliferation (MTS/XTT), migration/invasion, and glucose uptake assays. Co-immunoprecipitation can confirm disrupted GRB10-INSR/IGF1R binding. The model supports research in hepatocellular carcinoma, type 2 diabetes, growth disorders, and kinase inhibitor drug screening. For further information, contact Ascent Research.
