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Pseudoislet formation enhances gene expression, insulin secretion and cytoprotective mechanisms of clonal human insulin-secreting 1.1B4 cells

  • Signaling and cell physiology
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Abstract

We have studied the effects of cell communication on human beta cell function and resistance to cytotoxicity using the novel human insulin-secreting cell line 1.1B4 configured as monolayers and pseudoislets. Incubation with the incretin gut hormones GLP-1 and GIP caused dose-dependent stimulation of insulin secretion from 1.1B4 cell monolayers and pseudoislets. The secretory responses were 1.5–2.7-fold greater than monolayers. Cell viability (MTT), DNA damage (comet assay) and apoptosis (acridine orange/ethidium bromide staining) were investigated following 2-h exposure of 1.1B4 monolayers and pseudoislets to ninhydrin, H2O2, streptozotocin, glucose, palmitate or cocktails of proinflammatory cytokines. All agents tested decreased viability and increased DNA damage and apoptosis in both 1.1B4 monolayers and pseudoislets. However, pseudoislets exhibited significantly greater resistance to cytotoxicity (1.5–2.7-fold increases in LD50) and lower levels of DNA damage (1.3–3.4-fold differences in percentage tail DNA and olive tail moment) and apoptosis (1.3–1.5-fold difference) compared to monolayers. Measurement of gene expression by reverse-transcription, real-time PCR showed that genes involved with insulin secretion (INS, PDX1, PCSK1, PCSK2, GLP1R and GIPR), cell-cell communication (GJD2, GJA1 and CDH1) and antioxidant defence (SOD1, SOD2, GPX1 and CAT) were significantly upregulated in pseudoislets compared to monolayers, whilst the expression of proapoptotic genes (NOS2, MAPK8, MAPK10 and NFKB1) showed no significant differences. In summary, these data indicate cell-communication associated with three-dimensional islet architecture is important both for effective insulin secretion and for protection of human beta cells against cytotoxicity.

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Acknowledgments

This work was funded by the University of Ulster Research Strategic funding and the award of a University of Ulster Vice Chancellor’s research studentship to SV and a Northern Ireland Department of Employment and Learning research studentship to ADG.

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The authors declare no conflicts of interest associated with this manuscript.

Human rights and animal welfare statement

This article does not contain any studies with human participants or animals performed by any of the authors.

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    Authors and Affiliations

    1. SAAD Centre for Pharmacy and Diabetes, University of Ulster, Cromore Road, Coleraine, BT52 1SA, Northern Ireland, UK

      Alastair D. Green, Srividya Vasu, Neville H. McClenaghan & Peter R. Flatt

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    1. Alastair D. Green
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    2. Srividya Vasu
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    3. Neville H. McClenaghan
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    4. Peter R. Flatt
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    Correspondence to Srividya Vasu.

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    Alastair D. Green and Srividya Vasu contributed equally to this work.

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    Green, A.D., Vasu, S., McClenaghan, N.H. et al. Pseudoislet formation enhances gene expression, insulin secretion and cytoprotective mechanisms of clonal human insulin-secreting 1.1B4 cells. Pflugers Arch - Eur J Physiol 467, 2219–2228 (2015). https://doi.org/10.1007/s00424-014-1681-1

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