The AGGF1 Knockout CAL-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the CAL-27 human tongue squamous cell carcinoma line. These cells carry targeted disruptions in the AGGF1 gene, leading to loss of functional angiogenic factor AGGF1. The polyclonal format preserves genetic diversity, suitable for studying AGGF1-dependent phenotypes in a heterogeneous context. The cells are designed for investigating AGGF1-mediated angiogenic signaling and tumor-endothelial crosstalk in oral cancer.
CAL-27 is an adherent epithelial cell line from a 56-year-old male with tongue squamous cell carcinoma. It is a standard oral cancer model with tumorigenic properties and robust xenograft tumor formation, enabling gene function studies in head and neck cancer. The epithelial origin supports investigations into tumor cell?Cmatrix interactions and paracrine signaling.
AGGF1 encodes a secreted angiogenic factor that activates PI3K/AKT, MAPK/ERK, and JAK/STAT pathways to promote endothelial cell proliferation, migration, and tube formation. It is thought to bind integrin ??v??3, initiating signaling through PIK3CA, AKT1, mTOR, MAP2K1, MAPK1/3, and JAK2/STAT3. AGGF1 expression is regulated by HIF1A, NF-??B, and STAT3, linking it to hypoxic and inflammatory conditions. Downstream, AGGF1 induces phosphorylation of AKT1 (p-AKT), MAPK1/3 (p-ERK), and STAT3 (p-STAT3), and indirectly upregulates VEGFA. In the knockout cells, loss of AGGF1 secretion eliminates autocrine survival signaling and paracrine endothelial activation, attenuating these pathways and impairing angiogenic capacity. This results in reduced tumor cell proliferation and diminished endothelial tube formation.
In oral squamous cell carcinoma, AGGF1-driven angiogenesis is essential for tumor progression. This knockout model enables dissection of AGGF1’s contribution to tumor cell-autonomous growth and paracrine vascular regulation. Elimination of AGGF1 is expected to reduce proliferation, migration, and endothelial network formation, targeting signaling nodes mTOR, p-AKT, p-ERK, and p-STAT3. The polyclonal population allows assessment of phenotypic variability and signaling thresholds relevant to angiogenic competence. The model offers insights into the role of AGGF1 in tumor heterogeneity and oral cancer pathogenesis.
Western blotting and RT-qPCR confirm AGGF1 knockout and quantify downstream effectors like p-AKT, p-ERK, and p-STAT3. Endothelial tube formation assays with conditioned medium evaluate paracrine angiogenic potential, while MTT and transwell assays measure proliferation and migration. In vivo xenograft tumor models with CD31 immunohistochemistry assess microvessel density, and co-culture systems enable study of tumor?Cendothelial communication. Researchers focusing on AGGF1 as a therapeutic target in oral cancer or investigating angiogenic signaling in solid tumors will benefit from this model. For technical details, please contact Ascent Research.
