Cadherin Engagement Protects Human β-Cells from Apoptosis

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Abstract

The aim of this study was to assess the expression of different types of cadherins in human islets and their role in human β-cell apoptosis. Expression of E-, N-, and P-cadherins was studied by immunofluorescence on pancreas sections and islet cells, and by Western blotting on protein extracts of isolated islets and islet cells. The effects of specific cadherins on cell adhesion and apoptosis were studied using chimeric proteins containing functional E-, N-, or P-cadherin ectodomains fused to Fc fragment of Ig (E-cad/Fc, N-cad/Fc, and P-cad/Fc) and immobilized on glass substrate. β-Cells were identified by immunofluorescence for insulin and apoptotic cells by terminal deoxynucleotide transferase-mediated 2′-deoxyuridine, 5′-triphosphate nick-end labeling. By immunofluorescence, we showed that E- and N-, and not P-, cadherins were expressed at the surface of islet cells. By triple staining, we showed that E-cadherin was expressed at similar extent in β- and α-cells, whereas N-cadherin was preferentially expressed in β-cells. These results were confirmed by Western blot analysis using protein extracts from fluorescence-activated cell sorting-sorted β- and non-β-cells. Adhesion tests showed that the affinity of islet cells for E-cad/Fc and N-cad/Fc and not for P-cad/Fc was increased compared with control. By terminal deoxynucleotide transferase-mediated 2′-deoxyuridine, 5′-triphosphate nick-end labeling, we showed that the percentage of apoptotic cells was lower in aggregated β-cells compared with single β-cells and that attachment to E-cad/Fc and N-cad/Fc and not to P-cad/Fc decreased apoptosis of single β-cells compared with control. Our results show that at least E- and N-cadherins are expressed at the surface of human β-cells and that these adhesion molecules are involved in the maintenance of β-cell viability.

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