Description

The DKK1 Knockout NCI-N87 Cell Line is a CRISPR/Cas9-edited knockout cell line with disruption of the DKK1 gene in the human NCI-N87 gastric carcinoma cell line. This model abrogates Dickkopf-related protein 1 (DKK1) expression via CRISPR/Cas9-mediated gene disruption, providing a stable loss-of-function system for investigating Wnt/??-catenin signaling in a gastric adenocarcinoma background.

The parental NCI-N87 cell line originates from a liver metastasis of a well-differentiated human gastric adenocarcinoma. As a HER2-overexpressing gastric carcinoma epithelial cell line, NCI-N87 is widely used for studying gastric cancer biology and therapeutic responses, particularly to HER2-targeted agents. Its adherent epithelial growth and robust proliferative capacity support diverse experimental paradigms.

DKK1 is a secreted Wnt antagonist that binds LRP5/6 coreceptors and Kremen1/2, blocking Wnt ligand?Cinduced Frizzled?CLRP5/6 complex formation and inhibiting ??-catenin stabilization. DKK1 transcription is regulated by ??-catenin/TCF, p53, TGF-??, and the vitamin D receptor. Upon knockout, loss of DKK1-mediated LRP5/6 inhibition results in constitutive Wnt pathway activation: ??-catenin accumulates, translocates to the nucleus, and partners with TCF7/LEF1 to transactivate target genes including c-Myc, Cyclin D1, Axin2, and survivin (BIRC5). This disrupts feedback and crosstalk with PI3K/Akt and MAPK/ERK cascades.

In the NCI-N87 context, DKK1 knockout releases its tumor-suppressive constraints, enhancing ??-catenin-driven proliferation, survival, and metastatic potential. Coupled with the cell line??s HER2 overexpression, this model permits dissection of collaborative signaling between Wnt/??-catenin and growth factor pathways, reflecting the molecular landscape of gastric adenocarcinoma. It is thus ideally suited for mechanistic studies and preclinical evaluation of Wnt-directed therapies.

Applications include western blotting and immunofluorescence for ??-catenin localization, RT-qPCR analysis of Wnt target genes (AXIN2, MYC, CCND1), TOPFlash/FOPFlash reporter assays for TCF/LEF activity, and phenotypic assays for proliferation, apoptosis, migration, invasion, and colony formation. The line also enables drug sensitivity profiling with Wnt inhibitors and transcriptomic analysis via RNA-seq. For more information, please contact Ascent Research.