The IGF2BP3 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human hepatic adenocarcinoma cell line. They feature targeted disruption of the IGF2BP3 gene, resulting in a loss-of-function model for studying RNA-binding protein-mediated post-transcriptional regulation. This polyclonal population provides a heterogeneous pool of cells with diverse editing outcomes, suitable for bulk analyses and functional screens without clonal selection artifacts.
SK-HEP-1 is a widely used liver cancer cell line originally isolated from the ascitic fluid of a patient with hepatic adenocarcinoma. Despite its hepatic origin, it exhibits an endothelial-like phenotype, expressing factors such as CD34 and factor VIII-related antigen, and forms capillary-like structures in vitro. This unique dual nature makes it a valuable model for investigating cancer biology, particularly the interplay between epithelial and endothelial characteristics, angiogenesis, and metastasis. The cell line??s robust growth and extensive characterization in hepatocellular carcinoma research provide a reliable platform for functional genomics studies.
IGF2BP3 (IMP3) is an oncofetal RNA-binding protein that post-transcriptionally enhances the stability and translation of target mRNAs. It directly interacts with transcripts of oncogenic factors including IGF2, MYC, CD44, and MMP9 via specific RNA motifs. Upstream, IGF2BP3 is regulated by transcription factors MYC, CTNNB1 (??-catenin), and E2F1, and suppressed by miRNAs let-7, miR-145, and miR-200 family. It forms complexes with proteins such as IGF2BP1, IGF2BP2, ELAVL1 (HuR), eIF4E, and PABPC1 to enhance oncogenic translation. This activates IGF2-IGF1R-PI3K-AKT-mTOR and WNT3A-??-catenin pathways, driving EMT through SNAI1 and TWIST1.
In the SK-HEP-1 liver cancer background, IGF2BP3 knockout provides a powerful tool to dissect hepatocarcinogenesis and metastasis. These cells express endothelial markers and exhibit aggressive behavior partly due to IGF2BP3 stabilization of prometastatic mRNAs. Disruption reduces expression of targets like CD44, MMP9, and VIM, diminishing motility, invasion, and anoikis resistance. This model is relevant for studying post-transcriptional control in liver cancer progression, EMT, and cancer stem cell-like properties.
These polyclonal knockout cells are suitable for RNA-seq, quantitative proteomics, RNA immunoprecipitation, and RNA stability assays to assess gene expression changes and mRNA target validation. Functional assays include wound healing, transwell invasion, MTT, colony formation, and immunofluorescence to evaluate migration, proliferation, and EMT markers. In vivo models apply orthotopic or tail-vein xenografts for metastasis. Drug sensitivity screening can assess chemoresistance. For further information, please contact Ascent Research.
