The CD44 Knockout Huh-7 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 human hepatocellular carcinoma line. This loss-of-function model disrupts CD44, the primary receptor for hyaluronan, without clonal isolation, preserving population-level heterogeneity for studies requiring biological variation in pooled functional genomics or signaling analyses.
Huh-7 cells were originally isolated from a liver tumor of a 57-year-old Japanese male and are widely employed as a hepatocellular carcinoma model, retaining hepatocyte-like metabolic properties and tumorigenic capacity. This cell line supports robust gene editing and serves as a relevant host for probing hepatocarcinogenesis, viral hepatitis interactions, and liver-specific drug metabolism in the context of CD44 ablation.
CD44 is a transmembrane glycoprotein that mediates cell?Ccell and cell?Cmatrix adhesion and transduces signals from the extracellular matrix component hyaluronan. Upstream regulators including TNF-??, IL-1??, EGF, and TGF-?? drive CD44 expression through NF-??B, AP-1, and STAT3. Upon ligand binding, CD44 engages PI3K/AKT and MAPK/ERK cascades, activating downstream effectors such as AKT, ERK1/2, Rac1, and RhoA. This signaling promotes epithelial?Cmesenchymal transition (EMT), upregulates matrix metalloproteinases like MMP-9, and stabilizes ??-catenin, thereby enhancing cell migration, invasion, and stem cell marker expression (Nanog, Oct4).
In hepatocellular carcinoma, CD44 overexpression is associated with metastasis and poor prognosis. The Huh-7 CD44 knockout polyclonal cells allow dissection of hyaluronan-mediated adhesion, migration, and EMT under controlled conditions. This population is ideal for studying heterogeneous responses to cytokine stimulation and for evaluating inhibitors of PI3K/AKT or MAPK/ERK pathways. It also facilitates investigation of the cancer stem cell fraction and its contribution to chemoresistance and tumor relapse.
Applications include western blotting and flow cytometry for CD44 and EMT markers, migration/invasion assays, phospho-signaling analysis (p-AKT, p-ERK), co-immunoprecipitation with hyaluronan, RNA-seq, and drug sensitivity testing. The model supports research in cancer biology, metastasis, drug resistance, and tumor microenvironment studies in liver cancer. For technical assistance, please contact Ascent Research.
