The ATG2B Knouckout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting ATG2B in the human SK-HEP-1 cell line. This loss-of-function model enables investigation of ATG2B-dependent processes in autophagy and related pathways. The polyclonal format mitigates clonal variation and is suited for bulk-population biochemical assays and functional screens.
SK-HEP-1 is a human epithelial cell line isolated from ascitic fluid of a liver adenocarcinoma patient. It is extensively used in hepatocellular carcinoma research, providing a relevant model for studying tumor biology, metastasis, and drug response. Its adherent growth and established utility in autophagy studies make it apt for dissecting ATG2B??s role in liver cancer cell physiology.
ATG2B encodes a lipid transfer protein central to autophagosome biogenesis. It acts downstream of the ULK1 kinase complex and the PI3KC3 lipid kinase, which generates phosphatidylinositol 3-phosphate (PI3P) at endoplasmic reticulum subdomains. ATG2B is recruited to these sites through interaction with the PI3P-binding PROPPIN family proteins WIPI1 and WIPI2. Once localized, ATG2B forms a functional complex with ATG9A and transfers lipids from the ER to the expanding isolation membrane, facilitating phagophore expansion and autophagosome closure. This process is tightly regulated: mTORC1 suppresses autophagy under nutrient-rich conditions, while amino acid deprivation activates ULK1 to stimulate the pathway. Additionally, TFEB transcriptionally upregulates ATG2B expression upon lysosomal stress. ATG2B collaborates with LC3B for lipid conjugation; its disruption impairs LC3 lipidation and p62/SQSTM1 degradation, blocking autophagic flux and autolysosome maturation.
In SK-HEP-1 liver adenocarcinoma cells, ATG2B knockout provides a critical model to examine autophagy??s role in hepatocellular carcinoma progression. Autophagy is often dysregulated in liver cancer, contributing to cell survival, metabolic reprogramming, and resistance to chemotherapy. Loss of ATG2B disrupts lipid transfer during autophagosome formation, allowing researchers to investigate how defective lipid trafficking impacts cellular bioenergetics, proliferation, and drug sensitivity. This model is especially valuable for studying autophagy-related lipid metabolism in cancer and for evaluating how impaired autophagosome biogenesis sensitizes or protects tumor cells to therapeutic agents.
The ATG2B Knouckout SK-HEP-1 Polyclonal Cells support diverse applications in autophagy research, liver cancer biology, lipid metabolism, and drug resistance studies. Key assays include Western blotting to monitor LC3B lipidation and p62 accumulation, fluorescence microscopy to quantify LC3 puncta formation, and autophagy flux assays using inhibitors of lysosomal degradation. Functional measurements such as cell proliferation, migration, invasion, and drug sensitivity complement the autophagic readouts. Genotyping PCR confirms CRISPR/Cas9-mediated gene disruption. The polyclonal format is well-suited for large-scale screening and robust biochemical analyses. For additional details, please contact Ascent Research.
