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Id3 upregulates BrdU incorporation associated with a DNA damage response, not replication, in human pancreatic ?-cells.

ABSTRACT: Elucidating mechanisms of cell cycle control in normally quiescent human pancreatic ?-cells has the potential to impact regeneration strategies for diabetes. Previously we demonstrated that Id3, a repressor of basic Helix-Loop-Helix (bHLH) proteins, was sufficient to induce cell cycle entry in pancreatic duct cells, which are closely related to ?-cells developmentally. We hypothesized that Id3 might similarly induce cell cycle entry in primary human ?-cells. To test this directly, adult human ?-cells were transduced with adenovirus expressing Id3. Consistent with a replicative response, ?-cells exhibited BrdU incorporation. Further, Id3 potently repressed expression of the cyclin dependent kinase inhibitor p57 (Kip2 ) , a gene which is also silenced in a rare ?-cell hyperproliferative disorder in infants. Surprisingly however, BrdU positive ?-cells did not express the proliferation markers Ki67 and pHH3. Instead, BrdU uptake reflected a DNA damage response, as manifested by hydroxyurea incorporation, ?H2AX expression, and 53BP1 subcellular relocalization. The uncoupling of BrdU uptake from replication raises a cautionary note about interpreting studies relying solely upon BrdU incorporation as evidence of ?-cell proliferation. The data also establish that loss of p57 (Kip2) is not sufficient to induce cell cycle entry in adult ?-cells. Moreover, the differential responses to Id3 between duct and ?-cells reveal that ?-cells possess intrinsic resistance to cell cycle entry not common to all quiescent epithelial cells in the adult human pancreas. The data provide a much needed comparative model for investigating the molecular basis for this resistance in order to develop a strategy for improving replication competence in ?-cells.

SUBMITTER: Lee SH

PROVIDER: S-EPMC3329516 | biostudies-literature | 2011 Nov-Dec

REPOSITORIES: biostudies-literature

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Id3 upregulates BrdU incorporation associated with a DNA damage response, not replication, in human pancreatic β-cells.

Lee Seung-Hee SH Hao Ergeng E Levine Fred F Itkin-Ansari Pamela P

Islets 20111101 6

Elucidating mechanisms of cell cycle control in normally quiescent human pancreatic β-cells has the potential to impact regeneration strategies for diabetes. Previously we demonstrated that Id3, a repressor of basic Helix-Loop-Helix (bHLH) proteins, was sufficient to induce cell cycle entry in pancreatic duct cells, which are closely related to β-cells developmentally. We hypothesized that Id3 might similarly induce cell cycle entry in primary human β-cells. To test this directly, adult human β- ...[more]

PMID: 21964314

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Dataset Information

Id3 upregulates BrdU incorporation associated with a DNA damage response, not replication, in human pancreatic ?-cells.

Publications

Id3 upregulates BrdU incorporation associated with a DNA damage response, not replication, in human pancreatic β-cells.

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