The DNAJB12 Knockout SK-HEP-1 Polyclonal Cells product comprises a heterogeneous population of SK-HEP-1 cells engineered via CRISPR/Cas9-mediated disruption of the DNAJB12 gene, yielding a robust loss-of-function model. This polyclonal knockout pool enables examination of DNAJB12 functions in a hepatocellular carcinoma-derived cell background with endothelial-like properties, circumventing clonal selection biases while maintaining genetic diversity within the edited population.
SK-HEP-1 cells were originally isolated from the ascitic fluid of a patient with liver adenocarcinoma and concurrently exhibit features of both hepatic adenocarcinoma cells and liver sinusoidal endothelial cells. This dual character positions the cell line as a versatile platform for studying liver cancer biology, endothelial cell processes, and the interplay between tumor cells and the microenvironment, particularly in the context of protein homeostasis and signal transduction.
DNAJB12 is an endoplasmic reticulum (ER)-resident co-chaperone that recruits cytosolic HSPA8 (Hsp70) to the ER membrane via its J-domain, facilitating the extraction and retrotranslocation of misfolded proteins for ubiquitin-dependent proteasomal degradation in the ER-associated degradation (ERAD) pathway. It functions within a multi-protein complex that includes BAG6, VCP/p97, Derlin-1, and the E3 ligase HRD1, and is connected to the OS9 and Sel1L recognition machinery. Beyond ER quality control, DNAJB12 modulates endocytic trafficking and lysosomal degradation of the epidermal growth factor receptor (EGFR), thereby regulating downstream mitogenic signaling. Under ER stress, its expression is upregulated by the unfolded protein response (UPR) sensors ATF6, IRE1, and PERK, and by heat shock factor 1 (HSF1).
In SK-HEP-1 cells, which retain endothelial and adenocarcinoma characteristics, disruption of DNAJB12 offers a valuable tool to dissect how ERAD dysregulation and altered EGFR degradation influence liver cancer pathogenesis and endothelial-like behavior. This model permits investigation of crosstalk between the UPR and EGFR pathways, and may reveal synthetic vulnerabilities exploitable in hepatocellular carcinoma or other malignancies where ER stress and growth factor signaling converge.
Researchers can employ this polyclonal knockout cell pool in a variety of applications, including analyzing ERAD substrate accumulation and UPR activation via Western blotting and RT-qPCR (e.g., measuring CHOP, BiP, or ATF4), visualizing DNAJB12?CHsp70 complex formation by co-immunoprecipitation and immunofluorescence, and quantifying EGFR surface levels and internalization using flow cytometry. Additionally, the cells facilitate apoptosis and migration/invasion assays under pharmacological ER stress induction, enabling functional profiling of candidate therapeutics targeting ER proteostasis or EGFR-driven pathways in liver adenocarcinoma models. For further information or technical support, please contact Ascent Research.
