The ITGAM Knockout Ca Ski Polyclonal Cells are a CRISPR/Cas9-mediated ITGAM-knockout population in the Ca Ski cervical carcinoma line. This polyclonal pool contains diverse gene disruptions, providing a robust loss-of-function model for studying integrin alpha M (CD11b). Loss of ITGAM eliminates Mac-1 complex formation, enabling interrogation of adhesion and signaling functions without clonal bias. The polyclonal editing strategy yields a heterogeneous cell pool, capturing a range of loss-of-function phenotypes and avoiding artifacts of clonal selection.
Ca Ski cells, derived from an HPV16-positive cervical epidermoid carcinoma metastasis to the small intestine, are a standard model for HPV-driven cervical cancer. They exhibit p53 and Rb pathway dysregulation and are widely used for adhesion, invasion, and metastasis research. Introducing ITGAM knockout in this background facilitates analysis of integrin-mediated malignant phenotypes. The cell line’s retention of epithelial characteristics and HPV16 oncogenes makes it a physiologically relevant host for investigating how integrin ablation modulates tumor cell behavior.
ITGAM encodes CD11b, which pairs with CD18 to form Mac-1, a receptor for ICAM-1, iC3b, fibrinogen, and CD87. ITGAM is induced by PU.1 and C/EBPalpha in response to TNF, IL-1??, LPS, and CXCL12. Mac-1 activation triggers Syk and Pyk2 kinases, leading to phosphorylation of paxillin and activation of Rac1/RhoA, intersecting with PI3K-Akt, NF-??B, and ERK pathways. Knockout disrupts these axes, abolishing complement-mediated phagocytosis and adhesion signaling. Additionally, Mac-1 collaborates with CD14 and CD16 in innate immune recognition, but in the tumor context, aberrant ITGAM activation may drive pro-survival signals.
Although CD11b is not typical in cervical epithelium, ectopic expression may influence tumor-stroma interactions and metastasis. ITGAM knockout in Ca Ski cells enables dissection of integrin-driven migration, matrix remodeling, and transendothelial migration. It may clarify crosstalk with growth factor pathways and impact on drug sensitivity, offering a platform for mechanistic studies in metastasis and therapeutic resistance. Further, the knockout can be combined with reconstitution experiments to map functional domains of CD11b.
Applications include adhesion assays on ICAM-1/iC3b, transwell migration/invasion, phagocytosis, flow cytometry for CD11b, and immunoblotting/immunofluorescence for Syk, Pyk2, paxillin, and focal adhesions. This model supports research in tumor-immune interactions, integrin signaling, and metastasis inhibition. Researchers may also utilize these cells for high-content screening of integrin inhibitors or for co-culture studies with immune cells to assess tumor-host interplay. For custom services, contact Ascent Research.
