The TFF3 Knockout Hep-G2 Cell Line is a genetically defined human knockout model generated by CRISPR/Cas9-mediated disruption of the TFF3 gene in the Hep-G2 hepatocellular carcinoma cell line. This adherent cell product provides a stable loss-of-function platform for investigating trefoil factor 3 signaling in liver cancer biology. By ablating TFF3 expression, the cell line eliminates confounding adaptive responses, making it ideal for dissecting TFF3-dependent cellular processes.
Hep-G2 cells, originally derived from a liver hepatocellular carcinoma of a 15-year-old male, exhibit adherent epithelial morphology and retain key hepatic functions. They are widely used as a model for hepatocyte function, drug metabolism, and hepatocellular carcinoma research. The parental line responds to cytokines and growth factors, supporting signaling studies. The TFF3 knockout derivative maintains these characteristics while enabling specific interrogation of TFF3-mediated mechanisms in a relevant cancer context.
TFF3 is a secreted trefoil factor that promotes mucosal repair and is implicated in tumor progression. Its expression is upregulated by inflammatory cytokines IL-1??, TNF-??, and IL-6 via transcription factors STAT3, NF-??B, and HNF4A. Secreted TFF3 acts through receptors CXCR4 and EGFR, triggering PI3K/AKT and MAPK/ERK pathways, leading to phosphorylation of AKT and ERK1/2. It also activates NF-??B and STAT3, stabilizes ??-catenin, and stimulates mTOR, driving proliferation, survival, migration, and invasion. Interactions with mucins further support barrier function. Thus, TFF3 integrates multiple oncogenic signals relevant to hepatocellular carcinoma.
In Hep-G2 cells, TFF3 knockout abrogates autocrine/paracrine stimulation of PI3K/AKT, MAPK/ERK, and NF-??B cascades, resulting in reduced cell proliferation, impaired migration, attenuated invasion, and enhanced apoptosis sensitivity. This genetic ablation therefore provides a precise tool to study the contribution of TFF3 to liver cancer maintenance, epithelial-mesenchymal transition, and metastatic potential, and to evaluate pathway-specific consequences in a hepatocellular carcinoma background devoid of TFF3.
The cell line supports diverse research applications, including western blot analysis of TFF3 and downstream phospho-proteins (AKT, ERK), cell viability (MTT) and apoptosis (annexin V/PI) assays, wound healing and transwell invasion assays, qPCR profiling of EMT markers, and immunofluorescence for ??-catenin localization. It is also suitable for xenograft tumor models to assess tumor growth and metastasis. For technical inquiries or custom product requests, please contact Ascent Research.
