The HGFAC Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from HCT 116 colorectal carcinoma cells, in which the HGFAC gene has been disrupted. This knockout eliminates hepatocyte growth factor activator (HGFA), a serine protease essential for the proteolytic activation of pro-hepatocyte growth factor (pro-HGF). The polyclonal format provides a genetically heterogeneous loss-of-function model suitable for pooled functional studies.
The parental HCT 116 line is a human colorectal carcinoma epithelial cell model that carries an oncogenic KRAS G13D mutation, exhibits microsatellite instability (MSI), and retains wild-type p53. This molecular background makes it a representative system for studying colorectal adenocarcinoma biology, including dysregulated proliferation, epithelial signaling, and early metastatic processes. The cells are widely utilized in cancer research for pathway analysis and drug screening applications.
The HGFAC gene product is a serum-derived serine protease that cleaves inactive pro-HGF into bioactive HGF, the sole ligand for the c-Met receptor tyrosine kinase. HGF binding triggers c-Met autophosphorylation and recruitment of adaptors GRB2 and GAB1, which activate PI3K/AKT, RAS/ERK, and STAT3 signaling cascades, promoting cell proliferation, scattering, and survival. HGFA is activated by upstream proteases including thrombin, plasma kallikrein, and coagulation factor XIIa, and is inhibited by serpins. Consequently, knockout of HGFAC eliminates pro-HGF processing and silences c-Met signaling.
In the HCT 116 KRAS-mutant background, HGF/c-Met signaling cooperates with constitutive RAS/ERK activity to enhance tumor cell proliferation, migration, and invasion. By eliminating HGF maturation, this knockout model enables selective interrogation of the HGF/c-Met axis independently of other receptor tyrosine kinase inputs. The cells are thus a pertinent tool for dissecting the protease-dependent activation of HGF and its contribution to colorectal cancer progression and metastasis.
Typical applications encompass mechanistic studies of HGF/c-Met signaling in colorectal carcinoma, functional analyses of protease-mediated growth factor activation, and screening of compounds targeting the MET kinase or HGF activation. Representative assay methods include Western blotting for activated c-Met and HGF, BrdU proliferation assays, scratch wound and Transwell migration/invasion assays, phospho-Met ELISA, RNA-seq transcriptomics, and co-immunoprecipitation of signaling complexes. For further details or custom inquiries, please contact Ascent Research.
