The BTK Knockout CAL-27 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population of the CAL-27 human tongue squamous cell carcinoma line, featuring disruption of the BTK gene. This heterogeneous pool provides a loss-of-function model to study Bruton’s tyrosine kinase (BTK) in an epithelial cancer setting, enabling investigation of BTK-dependent signaling beyond traditional B-cell contexts.
The parental CAL-27 cell line is a widely used model of oral squamous cell carcinoma, originally derived from a human tongue lesion. These malignant epithelial cells exhibit aggressive features such as dysregulated proliferation, migration, and survival, making them highly relevant for head and neck cancer research. CAL-27 cells have been reported to express BTK, offering a unique platform to interrogate the kinase’s role in solid tumor biology, an area where BTK functions remain poorly understood.
BTK is a non-receptor tyrosine kinase that functions downstream of the B-cell receptor (BCR) complex, where it is activated by SRC family kinases LYN and SYK. Upon activation, BTK phosphorylates PLCG2, triggering calcium flux and second messenger production, which in turn activate PKC, MAPK/ERK, and NF-??B pathways. BTK also modulates the PI3K-AKT axis through interactions with BLNK, GAB1, and PIK3R1. In CAL-27 cells, BTK knockout abolishes these downstream signals, including phosphorylation of PLCG2, ERK, and AKT, thus providing a defined background to dissect BTK-mediated proliferation and survival mechanisms.
In head and neck squamous cell carcinoma, aberrant kinase signaling contributes to malignant progression. BTK expression in CAL-27 suggests that BCR-like signaling modules may be co-opted in epithelial tumors to sustain growth. Disruption of BTK in this model removes a central signaling node, potentially reducing cell viability and motility. This knockout cell population is therefore valuable for validating BTK as a therapeutic target in solid tumors and for assessing responses to BTK inhibitors like ibrutinib in non-hematologic malignancies.
Typical applications include western blotting and RT-qPCR for knockout confirmation, immunofluorescence for localization studies, and phospho-signaling analysis of key downstream effectors such as phospho-PLCG2, phospho-ERK, and phospho-AKT. Functional assays like MTS/CCK-8 proliferation, Annexin V apoptosis, and transwell migration/invasion assays allow comprehensive phenotypic profiling. Drug sensitivity screening with ibrutinib enables assessment of BTK-targeted therapy in an epithelial context. For technical support or custom requests, please contact Ascent Research.
