The GOLPH3L Knockout SK-HEP-1 Polyclonal Cells product provides a population of CRISPR/Cas9-edited polyclonal knockout cells targeting the GOLPH3L gene in the SK-HEP-1 human hepatic cancer line. This mixed knockout cell pool carries heterogeneous loss-of-function mutations generated by CRISPR/Cas9-mediated gene disruption, enabling studies of GOLPH3L function without the clonal artifacts associated with single-cell-derived lines. As a polyclonal knockout cell population, it represents a genetically diverse model that avoids confounding effects from clonal selection, making it suitable for experiments where population-level phenotypes are of primary interest.
The SK-HEP-1 host cell line is an immortalized, adherent epithelial cell line originally established from the ascites of a patient with liver adenocarcinoma. Widely employed as a model for hepatocellular carcinoma, SK-HEP-1 cells exhibit properties of malignant hepatic tumors, including uncontrolled proliferation and metastatic potential. Their ascitic origin provides a clinically relevant context for studying peritoneal dissemination and tumor?Cmicroenvironment interactions. The line??s robust growth and transfection amenability allow efficient CRISPR/Cas9 editing, and its annotation as a hepatocellular carcinoma model supports research into hepatic oncogenesis and therapeutic response.
The GOLPH3L gene encodes a Golgi phosphoprotein 3-like protein that acts as a phosphatidylinositol-4-phosphate (PI4P) effector on trans-Golgi membranes. By binding PI4P and coupling to the actin cytoskeleton via MYO18A, it drives Golgi membrane extension and vesicle budding, facilitating secretory cargo delivery. GOLPH3L functions downstream of PI3K/AKT signaling and growth factor receptors such as EGFR and IGFR, is regulated by mTORC2, and in turn promotes mTORC1-mediated phosphorylation of S6K1 and 4EBP1. Additional interacting factors include Arf1 GTPase and coatomer complex components (e.g., COPA). This network links Golgi membrane dynamics to anabolic signaling and neoplastic growth.
In the SK-HEP-1 hepatocellular carcinoma background, GOLPH3L knockout provides a powerful system to dissect its role in hepatic malignancy. Loss of GOLPH3L is expected to disrupt Golgi organization and secretory function, attenuate mTORC1 signaling, and impair actin-based processes, which may collectively reduce proliferation, migration, and invasion. This model enables investigation of how GOLPH3L-mediated trafficking supports oncogenic signaling networks including PI3K/AKT/mTOR. Moreover, the polyclonal knockout population allows assessment of tumor heterogeneity and the selective pressures that might arise upon drug therapy, particularly in the context of mTOR inhibitors.
Key applications include western blotting for phosphorylated S6K1/4EBP1, immunofluorescence of Golgi markers (GM130, giantin), RT-qPCR of GOLPH3L, MTT/BrdU proliferation assays, Transwell migration/invasion, co-immunoprecipitation of MYO18A, cell cycle analysis by flow cytometry, and drug sensitivity testing with mTOR inhibitors. These assays support comprehensive dissection of GOLPH3L function in hepatic tumor biology. For additional details, please contact Ascent Research.
