The HMGB1 Knockout KYSE-150 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the KYSE-150 human esophageal squamous cell carcinoma cell line. This product consists of a heterogeneous pool of cells with targeted disruption of the HMGB1 gene, offering a loss-of-function model for investigating HMGB1-dependent processes in esophageal cancer. The polyclonal format avoids clonal selection artifacts and reflects the genetic diversity of the knockout population, making it suitable for bulk functional analyses.
KYSE-150 is a poorly differentiated esophageal squamous cell carcinoma cell line originated from a 49-year-old Japanese male patient. As an epithelial-derived cancer model, it retains key features of esophageal squamous cell carcinoma, including aggressive growth and invasive potential. This line is widely used to study tumor progression, metastasis, and therapeutic responses, providing a relevant context for examining HMGB1’s role in malignancy.
HMGB1 is a multifunctional protein that operates dually as a nuclear chromatin regulator and an extracellular damage-associated molecular pattern (DAMP). In the nucleus, it binds DNA and nucleosomes, interacting with p53 and TFIIB to facilitate transcription, DNA repair, and nucleosome assembly. Upon release, HMGB1 acts as a DAMP ligand for receptors such as RAGE, TLR4, and TLR2, triggering downstream signaling via NF-??B, MAPK (ERK, p38, JNK), and PI3K/Akt pathways. Extracellular HMGB1 signaling is activated by upstream factors including TNF-??, IL-1??, hypoxia, and LPS, while downstream it promotes expression of pro-inflammatory cytokines (IL-6, IL-8, TNF-??), matrix metalloproteinase MMP-9, cyclin D1, and anti-apoptotic Bcl-2. HMGB1 also interacts with CXCR4 and Beclin1 to modulate autophagy and cell migration.
Knockout of HMGB1 in KYSE-150 cells eliminates its nuclear regulatory functions and extracellular DAMP activities, disrupting RAGE- and TLR4-mediated activation of NF-??B and AP-1 transcription factors. This impairs expression of MMP-9 and Bcl-2, thereby reducing proliferative, invasive, and survival capacities. The model enables detailed dissection of how HMGB1 integrates signals from TNF-??, IL-1??, and hypoxia to sustain malignant phenotypes through PI3K/Akt and MAPK cascades, and it provides a platform to study autophagy and immune evasion mechanisms in esophageal cancer.
Typical applications include western blotting, RT-qPCR, and ELISA to confirm HMGB1 knockout and quantify secreted protein, immunofluorescence for subcellular localization, and functional assays such as MTT proliferation, Transwell migration/invasion, and NF-??B luciferase reporters. Co-immunoprecipitation and phospho-kinase arrays enable mapping of altered protein interactions and signaling networks, while RNA-seq transcriptome analysis reveals global gene expression changes. This polyclonal knockout population is well-suited for validating HMGB1-targeted therapeutics, studying DAMP-mediated inflammation and immune evasion, and exploring autophagy modulation in esophageal cancer. For further information, technical support, or customized services, please contact Ascent Research.
