The ITGB3 Knockout Ca Ski Polyclonal Cells comprise a CRISPR/Cas9-mediated gene-disrupted polyclonal population of Ca Ski cells, targeting the ITGB3 locus that encodes the ??3 integrin subunit. This heterogeneous knockout pool abrogates functional ??3 integrin expression, preventing formation of ??V??3 and ??IIb??3 heterodimers. The product serves as a loss?of?function tool for interrogating integrin?dependent adhesion, migration, and signal transduction in a defined cervical carcinoma background.
The parental Ca Ski line is an adherent epithelial cell model derived from a human cervical epidermoid carcinoma. It maintains multiple integrated copies (60?C600) of the high?risk human papillomavirus type 16 (HPV?16) genome and stably expresses the viral E6 and E7 oncoproteins, which inactivate tumor suppressors p53 and Rb. This genotype drives sustained proliferation and genomic instability, rendering Ca Ski cells a widely employed platform for studying HPV?mediated carcinogenesis, host?virus interactions, and therapeutic responses in cervical cancer.
ITGB3 encodes integrin ??3, which heterodimerizes with ??V or ??IIb to form receptors for vitronectin, fibronectin, fibrinogen, and other RGD ligands. Ligand binding induces talin- and kindlin-dependent activation, triggering the FAK?Src complex to propagate downstream signals. This cascade bifurcates into the PI3K?AKT?mTOR survival pathway and the Ras?Raf?MEK?ERK proliferative axis, while also controlling Rho GTPases (Rac1, CDC42, RhoA) that regulate cytoskeletal dynamics and focal adhesion turnover. Upstream modulators include VEGF, TGF???, PDGF, FGF, ECM stiffness, osteopontin, and IL?1??; the integrin also associates laterally with VEGFR2, PDGFR, and MMP2 to coordinate adhesion with growth factor and proteolytic signaling.
In the Ca Ski context, knockout of ITGB3 abolishes ??3?containing integrin heterodimers, thereby eliminating adhesion to vitronectin and fibronectin and attenuating outside?in signaling. This manifests as reduced phosphorylation of FAK Tyr397, Src Tyr416, Akt Ser473, and ERK1/2, along with impaired cytoskeletal reorganization and Rho GTPase activity. The cells consequently show decreased migration and invasion, and heightened sensitivity to anoikis. Because Ca Ski cells express HPV?16 E6/E7, the knockout also provides a unique platform to examine whether ??3 integrins synergize with viral oncoproteins to sustain anchorage?independent growth and metastatic dissemination.
Researchers can employ these polyclonal knockout cells in a range of functional assays, including adhesion assays on vitronectin or fibronectin substrates, Boyden chamber migration and Matrigel invasion assays, phospho?specific Western blotting for FAK, Akt, and ERK, immunofluorescence visualization of focal adhesions and actin stress fibers, flow cytometric apoptosis quantification, Rho GTPase activation measurements, and xenograft tumorigenicity studies. These applications are particularly suited to investigations of cervical cancer metastasis, integrin?mediated drug resistance, angiogenesis, and HPV?host interactions. For additional product details, pricing, or a quotation, please contact Ascent Research.
