Inhibition of α5 integrin decreases PI3K activation and cell adhesion of human colon cancers

Abstract

BackgroundPhosphatidylinositol 3-kinase (PI3K), acting through the downstream kinase AKT, regulates a number of cellular processes such as cell growth and survival. Poorly differentiated colon cancers are characterized by increased integrin-mediated extracellular matrix interactions; disruption of this integrin-mediated adhesion leads to apoptosis and involves reduced PI3K activity. The purpose of this study was to determine: (1) the role of α5β1 integrin expression on PI3K activity, and (2) whether blockade of α5β1 integrin decreases colon cancer cell adhesion and increases apoptosis. MethodsHuman colon cancer cells were treated with anti-α5 integrin antibody, anti-β1 integrin antibody, the PI3K inhibitor LY294002 (20 μmol/L), or wortmannin (400 nmol/L). Expression and function of α5 were quantitated by FACS analysis and cell adhesion assay, respectively. DNA fragmentation was measured to assess apoptosis. Protein was extracted to determine phosphorylated AKT (pAKT) expression as a measure of PI3K activity. ResultsIncreased pAKT and α5 integrin expression were noted in the colon cancer cells; blockade of α5 integrin decreased α5 integrin and pAKT expression and decreased cell adhesion. Moreover, PI3K inhibition resulted in decreased α5 integrin expression. In contrast, treatment with anti-β1 integrin antibody produced no change in pAKT expression or cell number. ConclusionOur results are the first to show that blockade of cell surface α5 integrin expression decreases PI3K activity, inhibits colon cancer cell attachment, and induces apoptosis. These findings suggest that agents which selectively target α5 integrin subunit expression may enhance the effects of standard chemotherapeutic agents and provide novel adjuvant treatment for selected colon cancers.