The GPSM1 Knockout SK-HEP-1 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal population of SK-HEP-1 cells bearing a targeted disruption of the GPSM1 gene, which encodes the G-protein signaling modulator 1 (AGS3). This loss-of-function model enables precise interrogation of GPSM1-dependent mechanisms in a hepatocellular carcinoma background. The polyclonal format provides a heterogeneous knockout pool that captures diverse genetic alterations, facilitating robust functional studies without the clonal selection biases inherent to monoclonal lines.
Derived from the ascitic fluid of a patient with liver adenocarcinoma, the SK-HEP-1 cell line represents a well-characterized human hepatocellular carcinoma model with hepatic epithelial morphology. These cells exhibit anchorage-independent growth and tumorigenic properties in vivo, making them a relevant system for investigating liver cancer biology. The line retains key signaling networks found in hepatocarcinogenesis, including those governing proliferation, migration, and autophagy, thus offering a physiologically pertinent context for GPSM1 loss-of-function studies.
GPSM1 functions as a guanine nucleotide dissociation inhibitor (GDI) for G??i/o subunits, binding and stabilizing them in the inactive GDP-bound state. This activity positions GPSM1 as a critical modulator of G-protein coupled receptor (GPCR) signaling. Mechanistically, GPSM1 is regulated by upstream factors such as GPCR agonists and protein kinase A (PKA), and it interacts with a network of partners including G??i/o subunits, GPSM2 (LGN), nuclear mitotic apparatus protein (NuMA), dynein intermediate chain, and RhoGEF Lbc. It acts downstream of G??i to assemble the LGN/NuMA/dynein ternary complex required for mitotic spindle orientation during asymmetric cell division. Additionally, GPSM1 suppresses mTORC1 activity in a G??i-dependent manner, thereby promoting autophagy. Representative pathway components encompass GPCR?CG??i?CGPSM1/AGS3?CLGN?CNuMA?Cdynein (spindle positioning), GPCR?CG??i?CGPSM1?CmTORC1 inhibition?Cautophagy, and Wnt/Frizzled?CDishevelled?CGPSM1/LGN?Ccell polarity axes.
In the SK-HEP-1 hepatic adenocarcinoma context, GPSM1 knockout enables dissection of its contributions to cancer cell polarity, asymmetric division, and autophagy-dependent survival. Given hepatocellular carcinoma often exhibits dysregulated mTOR signaling and autophagy, this model is well-suited to evaluate how loss of GPSM1 alters these processes and influences tumorigenic traits. The cells also provide a platform for assessing the role of GPSM1-mediated G-protein regulation in migration, invasion, and drug sensitivity, particularly to agents such as sorafenib that target oncogenic kinase pathways. The model??s relevance extends to studying GPSM1-associated neurodevelopmental disorders through comparative analyses of cell polarity mechanisms.
Researchers can employ this knockout tool in a variety of experimental designs: western blotting for GPSM1, G??i, and autophagy markers (LC3, p62); RT-qPCR to confirm gene disruption; migration and invasion assays (wound healing, transwell); proliferation assays (MTT, BrdU); autophagy flux measurements using bafilomycin A1 treatment; confocal microscopy for spindle orientation; co-immunoprecipitation of LGN and NuMA; cAMP accumulation assays; transcriptomic profiling by RNA-seq; and drug sensitivity testing. The polyclonal nature ensures representation of multiple knockout genotypes, enhancing statistical power in pooled functional screens. For further technical specifications or ordering information, please contact Ascent Research.
