The ATG2A Knockout SK-HEP-1 Polyclonal Cells product comprises a polyclonal population of SK-HEP-1 cells subjected to CRISPR/Cas9-mediated disruption of the ATG2A gene, resulting in a heterogeneous pool of loss-of-function variants. This knockout model is provided as a mixed culture, not a clonal isolate, and is suitable for studying ATG2A-dependent processes in a human hepatocellular carcinoma background.
The SK-HEP-1 cell line, originally derived from the ascitic fluid of a patient with liver adenocarcinoma, exhibits epithelial morphology and serves as a widely accepted in vitro model for hepatocellular carcinoma research. These cells retain characteristics relevant to hepatic tumor biology, making them appropriate for investigating autophagy-related mechanisms in liver cancer pathogenesis, metabolic adaptation, and therapeutic response.
ATG2A encodes a key lipid transfer protein that directly shuttles lipids from the endoplasmic reticulum to the growing phagophore, a critical step for autophagosome membrane expansion and closure. Within the autophagy cascade, ATG2A functions downstream of the ULK1 initiation complex and mTORC1/AMPK nutrient-sensing pathways, and acts in concert with ATG9A and the WIPI proteins (WIPI1, WIPI2) to promote LC3 lipidation and subsequent cargo receptor degradation. Knockout of ATG2A disrupts this lipid delivery, leading to impaired autophagosome formation, accumulation of p62/SQSTM1, and reduced autophagic flux, as evidenced by diminished LC3B conversion and puncta formation. The protein also interacts with ATG13, RB1CC1, and ATG18A, linking it to the broader PI3KC3 complex and Atg8 conjugation machinery.
In the context of hepatocellular carcinoma, autophagy plays a dual role in tumor suppression and promotion, and ATG2A-mediated lipid transfer is pivotal for sustaining autophagic activity under metabolic stress or therapeutic challenge. The SK-HEP-1 knockout model enables dissection of ATG2A??s contribution to liver cancer cell survival, chemoresistance, and mitochondrial quality control. It also provides a platform to explore potential synthetic lethal interactions and validate autophagy-targeting compounds in a disease-relevant setting.
This product is intended for advanced mechanistic studies of autophagosome biogenesis, drug target validation, metabolic stress assays, and biomarker discovery. Representative analytical techniques include western blotting for LC3B and p62, immunofluorescence detection of LC3 puncta, autophagic flux quantification using lysosomal inhibitors, electron microscopy for ultrastructural analysis, and cell viability assessments under nutrient deprivation. For further information, researchers may contact Ascent Research.
