The HSPB8 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human hepatocellular carcinoma line. This product is engineered to disrupt the HSPB8 gene, encoding a small heat shock protein that functions as a molecular chaperone in the chaperone-assisted selective autophagy (CASA) pathway. The polyclonal pool ensures a heterogeneous mixture of loss-of-function alleles, providing a robust model for studying the collective effects of HSPB8 deficiency without clonal selection. The CRISPR/Cas9-mediated gene disruption generates a reliable loss-of-function model suitable for investigating the role of HSPB8 in protein quality control and stress responses.
The SK-HEP-1 cell line is a well-established human hepatic adenocarcinoma model originally derived from the ascites of a male patient with liver adenocarcinoma. These cells display epithelial morphology and retain features relevant to hepatocellular carcinoma research, including dysregulated growth signaling and susceptibility to proteotoxic stress. As a liver cancer model, SK-HEP-1 expresses hepatic markers and is widely used to study oncogenic pathways, drug metabolism, and tumor cell biology. The combination of HSPB8 knockout with this cancer background provides a unique system to examine intersections between tumorigenesis and proteostasis networks.
HSPB8 is a small heat shock protein that functions as a co-chaperone in the chaperone-assisted selective autophagy (CASA) pathway. It forms a complex with BAG3 and HSPA8/Hsc70 or HSPA1A/Hsp70, which recruits the ubiquitin ligase STUB1/CHIP to direct misfolded proteins to the autophagy receptor SQSTM1/p62 and autophagosomal LC3 for lysosomal degradation. This process is induced by heat shock factor 1 (HSF1) under cellular stress conditions such as heat shock or oxidative stress. Interacting factors including HSPB1 and DNAJB2 fine-tune this network, and loss of HSPB8 impairs clearance of aggregation-prone proteins like mutant SOD1 and huntingtin.
In the SK-HEP-1 liver adenocarcinoma line, HSPB8 knockout compromises autophagic flux and elevates susceptibility to proteotoxic stress, as cancer cells depend on the CASA pathway to manage misfolded protein burden during proliferation. Disruption of the HSPB8-BAG3-HSP70 axis leads to accumulation of ubiquitinated aggregates and may alter survival signaling, providing a system to study how hepatocellular carcinoma cells cope with proteostatic challenges. This model also facilitates exploration of therapeutic strategies targeting protein quality control in liver cancer.
This polyclonal knockout cell population is suitable for protein aggregation studies, autophagy research, and cancer stress response modeling. Key assays include Western blotting for LC3 and p62 to measure autophagic flux, immunofluorescence for aggregate detection, cell viability under stress conditions, co-immunoprecipitation of CASA components, and flow cytometry for apoptosis. Applications extend to drug screening for proteostasis modulators and investigating mechanisms underlying distal hereditary motor neuropathy type II and Charcot-Marie-Tooth disease type 2L. For additional information, please contact Ascent Research.
