The GOLIM4 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for loss-of-function studies of GOLIM4, a Golgi integral membrane protein. This heterogeneous pool of SK-HEP-1 cells carries disrupted GOLIM4 alleles, enabling investigation of Golgi-dependent glycosylation and trafficking without clonal selection. The CRISPR-mediated gene disruption abolishes GOLIM4 expression, allowing dissection of its role in glycoprotein maturation and secretion. This polyclonal format captures population-level variability, suitable for screening and pathway analysis in liver cancer models.
The SK-HEP-1 cell line, derived from ascites of a liver adenocarcinoma patient, displays a hybrid epithelial/endothelial phenotype. Widely employed as a hepatocellular carcinoma and liver sinusoidal endothelium model, these cells exhibit anchorage-independent growth and invasive capacity. Their endothelial-like features facilitate studies of vascular mimicry and cell?Cmatrix interactions, while their hepatic origin supports analysis of liver-specific glycosylation. This dual phenotype provides a relevant context for examining Golgi-mediated modifications impacting cancer behavior and microenvironmental communication.
GOLIM4 encodes a Golgi-resident transmembrane scaffold that interacts with matrix proteins GOLGA2 and GORASP1, and COPI components like COPB1. It functions in N- and O-glycan biosynthesis, facilitating glycosylation of integrins and growth factor receptors. Disruption of GOLIM4 impairs glycan processing of downstream targets EGFR and MET, altering receptor stability and signaling. GOLIM4 is also implicated in the Golgi stress response, potentially regulated by upstream stress pathways, and coordinates glycoprotein maturation with glycosyltransferases MGAT1, ST6GAL1, and B4GALT1, influencing adhesion, migration, and proliferation.
In the SK-HEP-1 hepatocellular carcinoma model, GOLIM4 knockout reveals the importance of Golgi-mediated glycosylation in tumor progression. Aberrant glycosylation drives altered cell adhesion, immune evasion, and metastasis. Loss of GOLIM4 is expected to disrupt the glycan landscape, affecting integrin-mediated attachment and growth factor receptor signaling critical for oncogenic transformation. This model enables investigation of how Golgi dysfunction contributes to hepatocellular carcinoma pathology, including epithelial-to-mesenchymal transition and sinusoidal dissemination, and may uncover glycan-dependent vulnerabilities.
Key applications include Western blot for knockout validation, immunofluorescence with GM130 for Golgi morphology, and lectin-based glycosylation profiling. qPCR monitors glyco-gene expression changes, while migration/invasion and MTS assays assess functional outcomes. Flow cytometry quantifies cell surface glycoproteins, revealing receptor presentation defects. These polyclonal knockout cells support screening and mechanistic studies in glycosylation-related disorders, prostate cancer, and liver cancer. For additional specifications or custom inquiries, contact Ascent Research.
