The DKK1 Knockout HGC-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HGC-27 human gastric adenocarcinoma cell line. This product provides a genetically disrupted DKK1 model for loss-of-function studies of canonical Wnt/??-catenin signaling. The polyclonal nature ensures a heterogeneous genetic background, avoiding clonal selection bias while enabling assessment of gene function across a mixed population. CRISPR/Cas9-mediated gene targeting disrupts DKK1 expression, facilitating investigation of downstream signaling alterations.
HGC-27 is a gastric epithelial cell line originally isolated from a lymph node metastasis of a gastric adenocarcinoma. It serves as a widely used in vitro model for poorly differentiated gastric carcinoma, enabling studies of gastric cancer biology, tumor progression, and metastasis. The cell line retains key signaling pathways, including Wnt/??-catenin, making it suitable for investigating DKK1 function in a gastric cancer context. Its metastatic origin allows examination of how DKK1 loss influences cell migration, invasion, and metastatic potential.
DKK1 encodes a secreted antagonist of canonical Wnt/??-catenin signaling. It binds to Wnt co-receptors LRP5/6 and the transmembrane Kremen1/2 proteins, blocking Wnt-Frizzled-LRP5/6 complex formation and promoting clathrin-mediated endocytosis of the receptor complex. This inhibits ??-catenin stabilization and nuclear translocation, leading to reduced transcription of Wnt target genes such as c-Myc, Cyclin D1, and AXIN2. DKK1 expression is regulated by upstream factors including Wnt ligands, p53, TGF-??, and the ??-catenin/TCF complex, which collectively modulate its context-dependent effects on proliferation, differentiation, and apoptosis.
Wnt/??-catenin pathway dysregulation is frequent in gastric cancer and contributes to progression and chemoresistance. HGC-27 cells, with endogenous DKK1, allow evaluation of DKK1 loss on Wnt signaling. Disruption of DKK1 relieves inhibition on LRP5/6, enhancing ??-catenin stabilization and transcriptional activity. This model enables dissection of DKK1’s role in proliferation, apoptosis, migration, and invasion in gastric adenocarcinoma, relevant for studies on Wnt-targeted therapies and metastasis, given the metastatic origin of the host cells.
Typical applications include western blotting for ??-catenin and target proteins, RT-qPCR for Wnt target genes, and TOP/FOP flash luciferase reporter assays to assess ??-catenin-driven transcription. Cell proliferation (MTT, BrdU), apoptosis (Annexin V/PI), and migration/invasion (Transwell) assays can be performed. Co-immunoprecipitation confirms the disrupted DKK1-LRP6 interaction. Beyond gastric cancer, this model is applicable to bone metabolism and Alzheimer??s disease research where Wnt regulation by DKK1 is implicated. For further information or to inquire about this product, please contact Ascent Research.
