ABSTRACT
Epithelial cells deprived of normal matrix contact through the appropriate integrin heterodimer undergo apoptosis; this phenomenon has been termed “anoikis,” the ancient Greek word meaning “homelessness.” Anoikis prevents shed epithelial cells from colonizing elsewhere. This process is important for normal development, and its loss can contribute significantly to tumor malignancy. This chapter discusses the stable gene transfer and apoptosis assay methods that work best with epithelial cells. It describes transient assays that are much faster but whose track record is less established. MDCK cells are a widely used epithelial cell model system. Coding sequences candidate anoikis-regulators are blunt-end ligated into the Pmel site of this plasmid, resulting in their being transcribed from the CMV promoter.
