Pro- and antiapoptotic proteins regulate apoptosis but do not protect against cytokine-mediated cytotoxicity in rat islets and β-cell lines

J. Jason Collier, Patrick T. Fueger, Hans E. Hohmeier, Christopher B. Newgard

Research output: Contribution to journalArticle

55 Citations (Scopus)

Abstract

Type 1 diabetes results from islet β-cell death and dysfunction induced by an autoimmune mechanism. Proinflammatory cytokines such as interleukin-1β and γ-interferon are mediators of this β-cell cytotoxicity, but the mechanism by which damage occurs is not well understood. In the current study, we present multiple lines of evidence supporting the conclusion that cytokine-induced killing of rat β-cells occurs predominantly by a nonapoptotic mechanism, including the following: 1) A rat β-cell line selected for resistance to cytokine-induced cytotoxicity (833/15) is equally sensitive to killing by the apoptosis-inducing agents camptothecin and etoposide as a cytokine-sensitive cell line (832/13). 2) Overexpression of a constitutively active form of the antiapoptotic protein kinase Akt1 in 832/13 cells provides significant protection against cell killing induced by camptothecin and etoposide but no protection against cytokine-mediated damage. 3) Small interfering RNA-mediated suppression of the proapoptotic protein Bax enhances viability of 832/13 cells upon exposure to the known apoptosis-inducing drugs but not the inflammatory cytokines. 4) Exposure of primary rat islets or 832/13 cells to the inflammatory cytokines causes cell death as evidenced by the release of adenylate kinase activity into the cell medium, with no attendant increase in caspase 3 activation or annexin V staining. In contrast, camptothecin- and etoposide-induced killing is associated with robust increases in caspase 3 activation and annexin V staining. 5) Camptothecin increases cellular ATP levels, whereas inflammatory cytokines lower ATP levels in both β-cell lines and primary islets. We conclude that proinflammatory cytokines cause β-cell cytotoxicity primarily through a nonapoptotic mechanism linked to a decline in ATP levels.

Original languageEnglish (US)
Pages (from-to)1398-1406
Number of pages9
JournalDiabetes
Volume55
Issue number5
DOIs
StatePublished - 2006
Externally publishedYes

Fingerprint

Islets of Langerhans
Apoptosis
Cytokines
Cell Line
Camptothecin
Proteins
Etoposide
Adenosine Triphosphate
Annexin A5
Caspase 3
Cell Death
Staining and Labeling
Adenylate Kinase
bcl-2-Associated X Protein
Cytoprotection
Type 1 Diabetes Mellitus
Interleukin-1
Protein Kinases
Interferons
Small Interfering RNA

Keywords

  • γ-IFN, γ-interferon
  • IL, interleukin
  • iNOS, inducible nitric oxide synthase
  • SiRNA, small interfering RNA

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

Cite this

Pro- and antiapoptotic proteins regulate apoptosis but do not protect against cytokine-mediated cytotoxicity in rat islets and β-cell lines. / Collier, J. Jason; Fueger, Patrick T.; Hohmeier, Hans E.; Newgard, Christopher B.

In: Diabetes, Vol. 55, No. 5, 2006, p. 1398-1406.

Research output: Contribution to journalArticle

Collier, J. Jason ; Fueger, Patrick T. ; Hohmeier, Hans E. ; Newgard, Christopher B. / Pro- and antiapoptotic proteins regulate apoptosis but do not protect against cytokine-mediated cytotoxicity in rat islets and β-cell lines. In: Diabetes. 2006 ; Vol. 55, No. 5. pp. 1398-1406.
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AU - Newgard, Christopher B.

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AB - Type 1 diabetes results from islet β-cell death and dysfunction induced by an autoimmune mechanism. Proinflammatory cytokines such as interleukin-1β and γ-interferon are mediators of this β-cell cytotoxicity, but the mechanism by which damage occurs is not well understood. In the current study, we present multiple lines of evidence supporting the conclusion that cytokine-induced killing of rat β-cells occurs predominantly by a nonapoptotic mechanism, including the following: 1) A rat β-cell line selected for resistance to cytokine-induced cytotoxicity (833/15) is equally sensitive to killing by the apoptosis-inducing agents camptothecin and etoposide as a cytokine-sensitive cell line (832/13). 2) Overexpression of a constitutively active form of the antiapoptotic protein kinase Akt1 in 832/13 cells provides significant protection against cell killing induced by camptothecin and etoposide but no protection against cytokine-mediated damage. 3) Small interfering RNA-mediated suppression of the proapoptotic protein Bax enhances viability of 832/13 cells upon exposure to the known apoptosis-inducing drugs but not the inflammatory cytokines. 4) Exposure of primary rat islets or 832/13 cells to the inflammatory cytokines causes cell death as evidenced by the release of adenylate kinase activity into the cell medium, with no attendant increase in caspase 3 activation or annexin V staining. In contrast, camptothecin- and etoposide-induced killing is associated with robust increases in caspase 3 activation and annexin V staining. 5) Camptothecin increases cellular ATP levels, whereas inflammatory cytokines lower ATP levels in both β-cell lines and primary islets. We conclude that proinflammatory cytokines cause β-cell cytotoxicity primarily through a nonapoptotic mechanism linked to a decline in ATP levels.

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