The ISOC2 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of human SK-HEP-1 hepatic adenocarcinoma cells with targeted disruption of the ISOC2 gene. This polyclonal knockout pool enables loss-of-function studies in a heterogeneous cellular background, avoiding clonal selection biases. Stable CRISPR/Cas9-mediated gene disruption ensures ISOC2 inactivation for robust functional analysis across multiple experimental settings, making it a versatile tool for liver cancer research.
The SK-HEP-1 parental line was derived from ascitic fluid of a liver adenocarcinoma patient and is widely used as a hepatocellular carcinoma (HCC) model. These cells exhibit epithelial morphology with endothelial-like properties, relevant for studying tumor progression, angiogenesis, and apoptosis. SK-HEP-1 retains intact p53 signaling, mitochondrial apoptosis pathways, and oxidative stress responses, providing a suitable and clinically relevant context for investigating ISOC2 function.
ISOC2 encodes a mitochondrial protein that plays a critical role in regulating oxidative stress, mitochondrial metabolism, and apoptosis. Transcriptionally activated by p53 and induced by oxidative stress, ISOC2 localizes to mitochondria and interacts directly with ANT2 (SLC25A5), the adenine nucleotide translocator. This interaction inhibits the opening of the mitochondrial permeability transition pore (mPTP), thereby preserving mitochondrial membrane potential and preventing the release of cytochrome c into the cytosol. Consequently, ISOC2 suppresses the activation of caspase-9 and caspase-3 and modulates the balance of Bcl-2 family proteins such as BAX, ultimately promoting cell survival. Upstream regulators include p53 and DNA damage signals, while downstream targets encompass ANT2, caspase-3, and BAX, with p21 also participating in the broader p53-mediated response.
In SK-HEP-1 cells, ISOC2 knockout eliminates a key pro-survival mechanism, sensitizing the polyclonal population to mitochondrial-mediated apoptosis and oxidative stress. This model allows in-depth investigation of ISOC2??s impact on HCC cell viability, reactive oxygen species handling, and endothelial-like properties, such as vascular mimicry. The heterogeneous knockout pool mirrors the genetic diversity of tumors, facilitating robust studies on cellular senescence induction, chemotherapeutic vulnerability, and the balance between apoptosis and survival signaling in liver cancer.
Functional assays compatible with this model include Western blotting for caspase-3, cleaved caspase-9, Bcl-2, BAX, and p53; JC-1 mitochondrial membrane potential measurement; DCFDA-based ROS quantification; annexin V/propidium iodide flow cytometry for apoptosis; MTT or colony formation viability assays; and co-immunoprecipitation to assess ISOC2-ANT2 interaction. Applications range from hepatocellular carcinoma pathogenesis and drug resistance studies to p53 signaling and oxidative stress research. For technical inquiries or ordering information, please contact Ascent Research.
