The EEF2K Knockout HGC-27 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HGC-27 human gastric carcinoma cell line, featuring targeted disruption of the EEF2K gene. This polyclonal format provides a heterogeneous loss-of-function model, circumventing clonal selection artifacts and enabling robust assessment of gene function across a mixed genetic background.
The HGC-27 host cell line is a highly tumorigenic epithelial line established from a metastatic lymph node of an undifferentiated human gastric adenocarcinoma. It is widely used to model gastric cancer pathogenesis, including invasive growth, metastasis, and chemoresistance, owing to its well-defined signaling pathways and rapid proliferation.
EEF2K encodes a calmodulin-dependent kinase that phosphorylates eEF2 at Thr56, inhibiting translational elongation. Its activity is stimulated by calcium/calmodulin and AMPK, while it is negatively regulated by mTORC1 through S6K. Downstream, phospho-eEF2 suppresses protein synthesis, promotes autophagy, and enhances cell survival under nutrient deprivation and hypoxia. In the autophagy pathway, EEF2K connects to ULK1, a critical inducer of autophagosome formation. EEF2K interacts with calmodulin, eEF2, mTORC1 complex, and HSP90, thus integrating signals from mTOR, AMPK, and autophagy pathways.
In gastric adenocarcinoma, EEF2K contributes to metabolic reprogramming and therapy resistance by coordinating translational control and autophagic flux, thereby facilitating tumor progression. The EEF2K Knockout HGC-27 Polyclonal Cells provide a relevant model to investigate EEF2K??s role in drug sensitivity, apoptosis, and stress adaptation. Its significance extends to breast and colorectal cancers and neurodegenerative conditions.
This knockout model enables western blotting for p-eEF2 (Thr56) and LC3-II to monitor kinase activity and autophagy, along with colony formation, apoptosis, migration/invasion, drug sensitivity, and metabolic Seahorse assays. RT-qPCR further assesses transcriptional changes. These applications facilitate detailed mechanistic studies of EEF2K in gastric cancer biology and drug discovery. For further details or ordering, please contact Ascent Research.
