The INPP5F Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the INPP5F gene in the human SK-HEP-1 hepatic adenocarcinoma cell line. This loss-of-function model, generated via CRISPR-mediated gene disruption, allows researchers to investigate INPP5F??s regulatory role in phosphoinositide metabolism and signal transduction. The polyclonal format provides a heterogeneous population reflective of CRISPR editing outcomes without clonal isolation, making it a versatile tool for cancer cell signaling studies.
The parental SK-HEP-1 cell line originates from ascitic fluid of a 52-year-old male with hepatic adenocarcinoma and is a standard tumorigenic hepatocellular carcinoma model. These adherent epithelial cells harbor active PI3K/AKT and mTOR pathways, facilitating investigations into oncogenic signaling, tumor progression, and metastatic mechanisms. Their well-characterized genetics and culture adaptability support a wide variety of gene perturbation experiments. SK-HEP-1 cells are particularly suited for studies linking phosphoinositide signaling to liver cancer aggressiveness.
INPP5F encodes a phosphoinositide 5-phosphatase that dephosphorylates PIP3 to generate PI(3,4)P2, thereby counteracting PI3K signaling and suppressing AKT activation. This enzyme functions downstream of receptor tyrosine kinases such as EGFR and the insulin receptor, limiting PIP3 accumulation and PDK1-mediated AKT phosphorylation. Consequently, it dampens downstream effectors including mTORC1, GSK3??, FOXO, and S6K. INPP5F localizes to endosomes and interacts with APPL1 and endosomal sorting components, implying spatially restricted signal termination. Loss of INPP5F results in sustained PIP3 elevation and chronic AKT activation.
In SK-HEP-1 hepatocellular carcinoma cells, loss of INPP5F elevates PIP3 levels and induces sustained AKT activity, which drives enhanced cell proliferation, resistance to apoptosis, and increased migratory and invasive capacity. These phenotypic changes mimic aggressive liver cancer traits and highlight the importance of INPP5F as a potential tumor suppressor in PI3K/AKT-driven malignancies. The model also permits exploration of downstream metabolic reprogramming events, as AKT signaling promotes anabolic metabolism, and crosstalk with mTORC1 amplifies growth signals.
These polyclonal knockout cells are suited for a range of assays. Western blotting for phospho-AKT (Ser473) and phospho-S6 probes pathway activation, while RT-qPCR confirms INPP5F disruption. Proliferation can be assessed via MTT or BrdU incorporation, colony formation, and apoptosis by annexin V or caspase-3/7 activity. Migration and invasion are measured in Boyden chamber or wound healing models. Drug sensitivity studies with PI3K inhibitors (e.g., LY294002, BKM120) and metabolic assays for glucose uptake are applicable. For further support, please contact Ascent Research.
