This product consists of CRISPR/Cas9-edited polyclonal knockout HGC-27 cells with targeted disruption of the CBLL1 gene, providing a loss-of-function model for studying the E3 ubiquitin ligase CBLL1 (Hakai) in a human gastric carcinoma background. The polyclonal population contains a heterogeneous mixture of knockout alleles, enabling robust assessment of gene function without clonal selection bias. These cells serve as a versatile platform for investigating CBLL1-dependent mechanisms in epithelial-mesenchymal transition (EMT), cell adhesion, and metastatic progression.
The HGC-27 cell line is derived from the lymph node metastasis of an undifferentiated gastric adenocarcinoma and represents a highly motile, invasive gastric cancer model. It displays mesenchymal features, making it particularly suitable for EMT research and for dissecting signaling pathways that govern tumor cell plasticity and metastasis. HGC-27 cells endogenously express key components of the E-cadherin regulatory network, including c-Src, ??-catenin, and CBLL1 itself, providing a physiologically relevant context for functional studies.
CBLL1 functions as an E3 ubiquitin-protein ligase that, upon phosphorylation by the c-Src kinase, specifically interacts with and ubiquitinates E-cadherin, targeting it for proteasomal degradation. This process disrupts adherens junctions and releases ??-catenin from the membrane, which can translocate to the nucleus to promote transcription of EMT-associated genes such as Snail and Vimentin. CBLL1 operates within a multimeric SCF-type complex containing Cullin-1, Skp1, and Rbx1, and its activity is further modulated by upstream signals from TGF-?? and integrin engagement. Additional downstream targets include Cortactin and p120-catenin, linking CBLL1 to cytoskeletal remodeling and focal adhesion dynamics.
In HGC-27 cells, CBLL1 knockout abrogates c-Src-mediated E-cadherin ubiquitination, stabilizing cell?Ccell contacts and potentially reversing mesenchymal traits. This model allows researchers to dissect the interplay between ubiquitin-mediated proteolysis and EMT in gastric carcinoma, explore c-Src/CBLL1/E-cadherin signaling axis, and evaluate the role of CBLL1 in metastatic dissemination. The polyclonal nature of the knockout population mimics heterogeneous tumor cell populations, enhancing translational relevance for drug response and phenotypic screening.
Key applications include E-cadherin ubiquitination assays, co-immunoprecipitation of CBLL1 with E-cadherin and c-Src, scratch wound healing and Transwell migration/invasion assays, Western blot analysis of EMT markers (e.g., ??-catenin, Snail, Vimentin), immunofluorescence for E-cadherin localization, and detection of phospho-CBLL1. These cells are valuable for substrate identification of ubiquitin ligases, anti-metastatic drug screening, and studies on cell adhesion dynamics. For further information or custom inquiries, please contact Ascent Research.
