CD14 Knockout Ca Ski Polyclonal Cells are a genetically engineered cell population originating from the Ca Ski human cervical carcinoma line, featuring CRISPR/Cas9-mediated disruption of the CD14 gene. This polyclonal knockout product provides a robust loss-of-function tool for investigating CD14-dependent pathways in an epithelial malignancy context.
The Ca Ski cell line is an adherent epithelial line derived from an epidermoid carcinoma metastasis in the small intestine. It stably harbors integrated human papillomavirus type 16 (HPV-16) genomes and expresses the E6 and E7 oncoproteins, thereby serving as a widely accepted model for HPV-associated cervical cancer. This cellular background is particularly suited for studying innate immune signaling within transformed epithelial cells.
CD14 encodes a glycosylphosphatidylinositol-anchored protein that functions as an essential coreceptor for bacterial lipopolysaccharide (LPS). CD14 interacts with LPS bound to LPS-binding protein (LBP) and then transfers LPS to the toll-like receptor 4 (TLR4)/myeloid differentiation factor 2 (MD-2) complex, initiating intracellular signaling. Downstream, MyD88, IRAK1, and TRAF6 are recruited, leading to activation of TAK1 and the IKK complex, which in turn phosphorylate and activate NF-kB. Concurrently, mitogen-activated protein kinases (MAPKs) including ERK, JNK, and p38 are stimulated. These cascades regulate transcription of proinflammatory mediators such as tumor necrosis factor (TNF), interleukin-6 (IL-6), and interleukin-1 beta (IL-1beta), and promote production of reactive oxygen species. CD14 expression is itself induced by stimuli like TNF-alpha, IL-1beta, interferon-gamma, and GM-CSF, and is controlled by transcription factors Sp1 and AP-1. In the absence of CD14, LPS-induced signaling is profoundly impaired, confirming its crucial role in innate immune activation.
In the Ca Ski model, CD14 knockout allows dissection of how cervical carcinoma cells respond to bacterial products through TLR4, and how such signals intersect with HPV oncoprotein-driven pathways. Given that NF-kB and MAPK cascades are often dysregulated in cancers and can be modulated by viral proteins, this system enables exploration of whether innate immune activation alters tumor cell proliferation, survival, or the inflammatory tumor microenvironment.
Researchers can utilize these polyclonal knockout cells for LPS dose-response experiments paired with NF-kB luciferase reporter assays, cytokine quantification via ELISA (e.g., TNF, IL-6), and western blot analysis of phospho-NF-kB and MAPK activation. Flow cytometry permits confirmation of CD14 surface ablation. The cells are appropriate for drug screening campaigns targeting the TLR4/CD14 axis and for studies examining the crosstalk between HPV E6/E7 and innate immune signaling. For further information, please contact Ascent Research.
