The IGF2R Knockout SK-HEP-1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human hepatic adenocarcinoma cell line. This product enables loss-of-function studies of the IGF2R gene (cation-independent mannose 6-phosphate receptor, CD222) in a heterogeneous cell background, allowing researchers to investigate gene function without clonal artifacts. The polyclonal format retains genetic diversity while ensuring robust IGF2R disruption across the population, making it suitable for pooled functional assays.
The parental SK-HEP-1 cell line was originally established from the ascitic fluid of a patient with liver adenocarcinoma and displays endothelial-like characteristics alongside its epithelial tumor origin. SK-HEP-1 cells are widely employed to model liver sinusoidal endothelial cells (LSECs) and the hepatic tumor microenvironment, providing a unique platform to study intercellular communication, angiogenesis, and cancer biology within the liver. The line??s dual identity offers a versatile context for investigating gene functions relevant to both tumor parenchyma and stromal interactions.
IGF2R encodes a type I transmembrane glycoprotein that serves as the cation-independent mannose 6-phosphate receptor (CI-MPR), primarily mediating the trafficking of M6P-tagged lysosomal acid hydrolases (e.g., cathepsin D/CTSD) from the trans-Golgi network to lysosomes via clathrin?CGGA adaptor complexes, and the endocytosis and lysosomal degradation of insulin-like growth factor 2 (IGF2). Loss of IGF2R therefore disrupts lysosomal enzyme delivery and IGF2 clearance, leading to lysosomal dysfunction and sustained IGF2?CIGF1R signaling. The receptor is transcriptionally regulated by WT1 and subject to epigenetic silencing at the imprinted IGF2R locus. Downstream, IGF2R functions as a tumor suppressor by restraining IGF1R/AKT/mTOR pathway activation; its absence permits unchecked phosphorylation of AKT and downstream effectors. Additionally, IGF2R interacts with the urokinase-type plasminogen activator receptor (uPAR) and can activate latent TGF-??, linking lysosomal trafficking to growth factor activation. These interlaced signals position IGF2R at a regulatory node intersecting nutrient sensing, lysosomal homeostasis, and growth control.
In the SK-HEP-1 background, IGF2R knockout creates a relevant model for dissecting tumor suppressor mechanisms in hepatocellular carcinoma (HCC) and liver endothelial biology. The loss of IGF2R??s canonical IGF2 clearance function results in autocrine/paracrine IGF2 overstimulation of IGF1R, driving IRS1/PI3K/AKT/mTOR proliferative signaling??a pathway frequently dysregulated in HCC. Concurrently, impaired lysosomal enzyme transport compromises functional lysosomal activity, potentially affecting autophagic flux and cellular metabolism, which are critical in the nutrient-stressed tumor microenvironment. Because SK-HEP-1 cells exhibit both endothelial and tumor cell features, this knockout model can be used to study how IGF2R loss influences angiogenesis, cell?Ccell adhesion, and the reciprocal communication between tumor cells and liver sinusoidal endothelium.
Researchers can employ this polyclonal knockout population for diverse applications. For tumor suppressor studies, it supports western blot and RT-qPCR validation of IGF2R ablation, phospho-AKT monitoring following IGF2 stimulation, and MTS cell proliferation assays. Lysosomal trafficking defects can be assessed via LAMP1 immunofluorescence and enzyme activity assays. The cells are also useful for M6P and IGF2 binding assays, co-immunoprecipitation, and RNA-seq profiling. Additionally, this knockout platform enables screening of IGF1R and mTOR inhibitors in a liver cancer context. For further technical specifications, please contact Ascent Research.
