The CD109 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human hepatocellular carcinoma cell line Huh-7. This product features targeted disruption of the CD109 gene, generating a loss-of-function model that enables investigation of CD109-dependent molecular mechanisms without single-cell cloning.
The Huh-7 cell line is a well-differentiated hepatocellular carcinoma model originally isolated from a 57-year-old male liver tumor. Exhibiting epithelial morphology, it is extensively used in liver cancer research, including hepatitis C virus (HCV) replication, tumor progression, and therapeutic testing, providing a clinically relevant context for studying CD109 function.
CD109 encodes a glycosylphosphatidylinositol (GPI)-anchored glycoprotein that functions as a transforming growth factor-beta (TGF-beta) co-receptor. It forms complexes with TGFBR1, TGFBR2, and CD44 to promote receptor internalization and degradation, thereby attenuating SMAD2 and SMAD3 phosphorylation. CD109 also activates JAK/STAT3 signaling independently, leading to STAT3 phosphorylation and downstream transcriptional effects. Key upstream regulators include TGFB1, TGFB2, TGFB3, EGF, SMAD3, and STAT3, while downstream targets encompass phospho-SMAD2, phospho-SMAD3, STAT3, p21, c-MYC, and VEGFA. CD109 physically interacts with EGFR, SMAD7, and the TGF-beta receptors, integrating multiple signaling pathways.
In the Huh-7 hepatocellular carcinoma model, CD109 knockout disrupts the balance between canonical TGF-beta/SMAD and non-canonical JAK/STAT3 pathways. Loss of CD109 is anticipated to enhance TGF-beta-induced SMAD2/3 phosphorylation and reduce STAT3 activation, potentially affecting cell proliferation, migration, invasion, and epithelial-mesenchymal transition. This knockout model enables dissection of CD109’s role in liver tumor progression, drug resistance, and cross-talk with growth factor signaling, including EGF/EGFR and PI3K/AKT pathways, thus identifying CD109-dependent vulnerabilities.
Researchers can utilize these CD109 knockout Huh-7 cells in western blot analysis of phospho-SMAD2/3, RT-qPCR of TGF-beta target genes, transwell migration and invasion assays, immunofluorescence localization, and flow cytometric assessment of surface CD109. TGF-beta-responsive luciferase and STAT3 transcriptional reporter assays enable pathway-specific readouts. Applications include mechanism-of-action studies, drug screening, and therapeutic target validation. For further technical details or custom services, please contact Ascent Research.
