Protein Kinase C-ζ Activation Markedly Enhances β-Cell Proliferation: An Essential Role in Growth Factor-Mediated β-Cell Mitogenesis.

Abstract

Abstract Diabetes results from a deficiency of functional β-cells. Previous studies have identified hepatocyte growth factor (HGF) and parathyroid hormone-related protein (PTHrP) as two potent β-cell mitogens.

Objectives: To determine: (i) whether HGF and PTHrP have additive/synergistic effects on β-cell growth and proliferation; (ii) the signaling pathways through which these growth factors mediate β-cell mitogenesis; and (iii) whether activation of this/these signaling pathway(s) enhances human β-cell replication.

Research Design and Methods: We generated and phenotypically analyzed doubly transgenic (DT) mice overexpressing PTHrP and HGF in the β-cell. INS-1 and primary mouse and human islet cells were used to identify mitogenic signaling pathways activated by HGF and/or PTHrP.

Results: Combined overexpression of HGF and PTHrP in the β-cell of DT mice did not result in additive/synergistic effects on β-cell growth and proliferation, suggesting potential cross-talk between signaling pathways activated by both growth factors. Examination of these signaling pathways in INS-1 cells revealed atypical protein kinase C (PKC) as novel intracellular target activated by both HGF and PTHrP in β-cells. Knock-down of PKCζ, but not PKCι/λ, expression using specific siRNAs blocked growth factor-induced INS-1 cell proliferation. Furthermore, adenovirus-mediated delivery of kinase-dead PKCζ completely inhibited β-cell proliferation in primary islet cells overexpressing PTHrP and/or HGF. Finally, adenovirus-mediated delivery of constitutively active PKCζ in mouse and human primary islet cells significantly enhanced β-cell proliferation.

Conclusions: PKCζ is essential for PTHrP and HGF-induced β-cell proliferation. PKCζ activation could be useful in therapeutic strategies for expanding β-cell mass in vitro and in vivo.