Phosphoinositide 3-kinases (PI3-kinases) are a family of enzymes that 3'-phosphorylate the inositol head group of membrane phosphoinositides. They are subdivided into three major classes (I, II and III) based on their structural homology, regulation and substrate specificity. It is now becoming clear that PI3-kinase isoforms are subject to differential regulation and may play distinct roles within the cell. PI3-kinases and their second messenger lipid products have been implicated in a plethora of cellular responses with increasing evidence for involvement in the pathogenesis of human diseases. The future development of specific PI3-kinase isoform inhibitors may offer therapeutic benefit in a broad range of clinical settings, related to cancer, inflammatory and immunological diseases.