The DNAJB4 Knockout HGC-27 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HGC-27 human gastric adenocarcinoma cell line. This polyclonal model, generated via targeted disruption of the DNAJB4 gene, provides a genetically heterogeneous pool of cells with loss-of-function mutations, enabling robust analysis of DNAJB4-dependent processes in a cancer-relevant background. The product retains the mixed clonal composition inherent to polyclonal editing, allowing assessment of phenotypic consistency without single-cell clone selection. It is supplied as a cryopreserved vial ready for direct culture and downstream applications.
HGC-27 is a poorly differentiated human gastric adenocarcinoma cell line originally established from a lymph node metastasis of an intestinal-type gastric carcinoma in a male patient. This cell line is widely utilized as a model for intestinal-type gastric cancer, exhibiting aggressive growth and molecular features of advanced disease. HGC-27 cells harbor alterations involving PI3K/Akt activation and p53 pathway dysregulation, making them especially suitable for investigating tumor suppressor mechanisms and oncogenic signaling. Their metastatic origin adds translational relevance for studies of invasion and progression.
DNAJB4 encodes an Hsp40/DnaJ co-chaperone that stimulates the ATPase activity of Hsp70 (HSPA1A/HSPA1B), facilitating protein folding, degradation, and stress responses. This co-chaperone functions as a tumor suppressor by stabilizing p53 and suppressing Akt signaling. Mechanistically, DNAJB4 interacts with Hsp70 and the ubiquitin ligase CHIP (STUB1) to manage client proteins. Its expression is activated by HSF1 under stress and regulated by p53. Knockout of DNAJB4 disrupts Hsp70-mediated proteostasis, leading to decreased p53 stability and enhanced Akt phosphorylation, shifting signaling toward proliferation and survival.
In the HGC-27 gastric cancer model, DNAJB4 ablation exacerbates tumorigenic properties by reinforcing oncogenic networks. Loss of co-chaperone function cripples the Hsp70 cycle, impairing p53’s pro-apoptotic and growth-suppressive activities while unleashing Akt-PI3K-mTOR signaling. This dual effect promotes increased cell proliferation, apoptosis resistance, and enhanced migratory and invasive capacities. Consequently, the knockout polyclonal cells recapitulate aggressive gastric adenocarcinoma features, offering a pathophysiologically relevant platform for dissecting chaperone-tumor suppression interplay.
These cells are suited for a wide array of research applications, including functional genomics of tumor suppressors, chaperone-mediated protein quality control, and signaling pathway validation. Typical assays include western blotting for DNAJB4, p53, Akt, and phospho-Akt; proliferation assays (MTT/CCK-8); apoptosis detection (Annexin V/PI); colony formation; and Transwell migration/invasion experiments. Co-immunoprecipitation can probe Hsp70 interactions. The model also facilitates drug sensitivity profiling, particularly for compounds targeting the p53?CMDM2?Cp21 axis or Akt signaling. For technical specifications or custom inquiries, contact Ascent Research.
