Pdx-1 Modulates Histone H4 Acetylation and Insulin Gene Expression in Terminally Differentiated α-TC-1 Cells

Abstract

Islet-associated transcription factors play a critical role in regulating pancreatic endocrine cell differentiation and islet hormone gene expression. Both alpha- and beta-cells differentiate from a common endocrine precursor cell. Therefore, it is important to reveal the differential gene expression profiles between alpha- and beta-cells that can direct their terminal cell fates. alpha-TC-1 and beta-TC-1 are 2 terminally differentiated islet tumor cell lines derived from islets transformed by promoter-specific driven SV40 T antigen overexpression. In this study, we demonstrated that Pdx-1 is a potent transcriptional regulator of endogenous insulin gene expression in alpha-TC-1 cells. Reverse transcriptase-polymerase chain reaction and chromatin immunoprecipitation assays were used to analyze gene expression patterns and chromatin modifications in the insulin promoter. Differential transcription factor expression profiles of alpha-TC-1 and beta-TC-1 cells revealed that INSM-1 and Pdx-1 transcription factors were expressed exclusively in beta-TC-1 cells. Overexpression of Ad-Pdx-1 in alpha-TC-1 cells induced insulin gene expression. Chromatin immunoprecipitation assays in Ad-Pdx-1 alpha-TC-1 cells demonstrated Pdx-1 occupancy and the hyperacetylation of histone H4 in the insulin promoter region. Collectively, these experiments revealed that Pdx-1 is a potent transcriptional regulator that is capable of modulating histone H4 acetylation and activates the insulin gene in a terminally differentiated glucagonoma cell line.