The IRS2 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the human SK-HEP-1 hepatic adenocarcinoma cell line, featuring targeted disruption of the IRS2 gene. This product provides a heterogeneous loss-of-function model that avoids clonal selection artifacts, making it suitable for bulk population studies of insulin and IGF-1 signaling. The polyclonal format allows for robust analysis of IRS2-dependent processes without the confounding effects of single-clone characteristics, and it can be used in a variety of metabolic and oncogenic signaling investigations.
SK-HEP-1 is an epithelial-like cell line originally isolated from the ascitic fluid of a patient with liver adenocarcinoma. It is widely employed as a model system for hepatocellular carcinoma, exhibiting properties relevant to liver cancer progression, metastasis, and drug response. The cells are amenable to genetic manipulation and provide a relevant hepatic background for studying the interplay between insulin signaling and cancer metabolism. Their adenocarcinoma origin makes them particularly valuable for exploring the role of metabolic regulators in liver tumor biology.
IRS2 (Insulin Receptor Substrate 2) is a key adaptor protein that mediates signal transduction downstream of activated insulin receptor (INSR) and insulin-like growth factor 1 receptor (IGF1R), as well as interleukin-4 (IL-4) stimulation. Following tyrosine phosphorylation by the receptors, IRS2 recruits SH2 domain-containing proteins including the p85 regulatory subunit of PI3K (PIK3R1) and the GRB2-SOS complex. This recruitment activates PI3K, leading to downstream signaling through AKT, GSK3??, FOXO1, mTOR, and S6K, which collectively promote glucose uptake, glycogen synthesis, cell growth, and survival. IRS2 thus serves as a central node linking metabolic and mitogenic pathways, with its disruption having profound effects on cellular homeostasis.
In the SK-HEP-1 liver adenocarcinoma context, knockout of IRS2 ablates critical insulin and IGF-1 signaling, resulting in impaired metabolic and proliferative responses that mimic features of hepatic insulin resistance and altered cancer metabolism. This model enables dissection of the specific contributions of IRS2 to hepatocarcinoma cell growth, glucose utilization, and sensitivity to pharmacological PI3K pathway inhibitors. It also provides a platform to investigate crosstalk between insulin resistance and cancer progression, which is relevant to obesity-associated hepatocellular carcinoma and type 2 diabetes.
Researchers can employ these polyclonal knockout cells in a broad range of experimental applications, including metabolic disease modeling, diabetes research, insulin signaling pathway dissection, and cancer metabolism studies. Representative assays include Western blotting for IRS2 and phospho-AKT, glucose uptake measurements, cell proliferation and viability assays, PI3K inhibitor sensitivity profiling, insulin stimulation experiments followed by phospho-signaling analysis, and RT-qPCR for downstream targets such as FOXO1 and S6K. The polyclonal nature supports high-throughput and long-term studies. For additional details, contact Ascent Research.
