The ASPSCR1 Knockout SK-HEP-1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the human ASPSCR1 gene in SK-HEP-1 cells. This heterogeneous polyclonal pool offers a loss-of-function system without clonal selection, facilitating studies on ASPSCR1 function in a genetically diverse background. The product serves as a versatile model to investigate ASPSCR1-dependent mechanisms in liver adenocarcinoma cells.
SK-HEP-1 is a human hepatic adenocarcinoma cell line derived from ascitic fluid, exhibiting adherent epithelial morphology and serving as an established model for hepatocellular carcinoma (HCC). Its hepatic origin and tumorigenic properties render it suitable for exploring liver-specific insulin signaling, glucose metabolism, and oncogenic pathways, providing a relevant cellular context for studying metabolic dysfunction in cancer.
ASPSCR1 tethers GLUT4 glucose transporters intracellularly via UBX domain interaction with p97/VCP and ubiquitinated cargo. Insulin-activated PI3K/AKT signaling phosphorylates ASPSCR1, releasing GLUT4 for plasma membrane translocation and glucose uptake. Additionally, ASPSCR1 engages in autophagy and ubiquitin-proteasome system function, interacting with ATG5 and forming a pathological TFE3 fusion in alveolar soft part sarcoma. Knockout likely causes GLUT4 surface accumulation and disrupts proteostasis and autophagy.
In SK-HEP-1 HCC cells, ASPSCR1 knockout enables dissection of insulin-regulated GLUT4 trafficking and its impact on glucose homeostasis, relevant to insulin resistance and metabolic syndrome. The model allows examination of fusion-independent ASPSCR1 roles in autophagy and protein degradation, which may influence cancer cell proliferation and migration. It provides a platform to study how GLUT4 dysregulation contributes to hepatocellular carcinoma metabolism.
Key applications include GLUT4 localization by immunofluorescence, glucose uptake assays, insulin stimulation with phospho-AKT western blot, p97/VCP co-immunoprecipitation, and autophagy flux analysis. Proliferation and Transwell migration assays facilitate functional readouts, while therapeutic screening can target GLUT4 trafficking or insulin sensitization. This knockout population supports target validation and mechanistic studies in liver cancer and metabolic disease. For further information, please contact Ascent Research.
