The DIXDC1 Knockout HGC-27 Polyclonal Cells constitute a polyclonal population of HGC-27 human gastric adenocarcinoma cells subjected to CRISPR/Cas9-mediated disruption of the DIXDC1 gene. This knockout model ablates DIXDC1 expression, enabling functional studies of this scaffold protein in gastric cancer cell biology.
The parental HGC-27 line derives from lymph node metastasis of a gastric adenocarcinoma and harbors a TP53 tumor suppressor mutation, recapitulating key genetic features of metastatic gastric cancer. These epithelial cells are widely employed as a model for gastric carcinoma metastasis and invasive disease, offering a clinically relevant context for studying pro-metastatic mechanisms.
DIXDC1 (Dishevelled-Axin Domain Containing 1) functions as a scaffold that positively regulates canonical Wnt/??-catenin signaling. It directly interacts with Dishevelled 2 (DVL2), Axin1, and LRP6 to stabilize ??-catenin and enhance TCF/LEF-mediated transcription of target genes such as CCND1 (cyclin D1) and MYC, which drive cell proliferation and migration. Beyond Wnt pathway engagement, DIXDC1 associates with PI3K and the serine/threonine kinase MARK1 (microtubule affinity-regulating kinase 1)/Par-1. By inhibiting MARK1 kinase activity, DIXDC1 prevents pathological phosphorylation of tau and concomitant microtubule destabilization, thereby preserving microtubule integrity and facilitating polarized cell motility. Additional interacting partners, including 14-3-3, further integrate DIXDC1 into signaling networks that coordinate cytoskeletal reorganization with transcriptional outputs. Upstream, DIXDC1 is activated by Wnt ligands such as Wnt3a and bridges the receptor complex to downstream effectors, establishing a link between extracellular cues and both ??-catenin-dependent gene expression and microtubule dynamics.
In the HGC-27 background, loss of DIXDC1 is expected to attenuate Wnt/??-catenin transcriptional activity and sensitize microtubules to depolymerization, impairing the proliferative and migratory capacity that characterizes metastatic gastric adenocarcinoma cells. The TP53 mutation present in this line further mirrors the genetic landscape of advanced gastric tumors, enhancing the translational relevance of DIXDC1 Loss in studying tumor progression under compromised tumor-suppressor function. Thus, these knockout cells provide a physiologically pertinent system for dissecting how DIXDC1 contributes to the metastatic phenotype and for interrogating its functional interactions with p53 status.
This knockout model supports a diverse array of experimental workflows. Researchers can assess alterations in cell proliferation via MTT or CCK-8 assays, evaluate migration and invasion through wound-healing scratch and transwell platforms, and quantify Wnt pathway activity using TOP/FOP flash luciferase reporters. Molecular profiling by Western blotting for DIXDC1, ??-catenin, phospho-MARK1, and downstream targets, as well as RT-qPCR for CCND1 and MYC, permits robust validation of pathway disruption. Immunofluorescence staining of microtubules reveals cytoskeletal consequences of MARK1 derepression. Co-immunoprecipitation assays enable mapping of DIXDC1 interactomes in the native gastric cancer milieu. Applications extend to biomarker discovery for gastric cancer, screening of Wnt or PI3K/AKT inhibitors, and functional genomics studies of DIXDC1-dependent signaling. For further technical information or to arrange custom bulk orders, please contact Ascent Research.
