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Cited by 100Pancreatic function and diabetesMicroRNA in disease regulationDiabetes and associated disorders

MicroRNA 21 targets BCL2 mRNA to increase apoptosis in rat and human beta cells

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Sims, Emily K., Lakhter, Alexander J., Anderson-Baucum, Emily, Kono, Tatsuyoshi, Tong, Xin, Evans-Molina, Carmella·2017·Diabetologia
DOI 10.1007/s00125-017-4237-zPMC PMC5425307
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Abstract

Aims/hypothesis The role of beta cell microRNA (miR)-21 in the pathophysiology of type 1 diabetes has been controversial. Here, we sought to define the context of beta cell miR-21 upregulation in type 1 diabetes and the phenotype of beta cell miR-21 overexpression through target identification. Methods Islets were isolated from NOD mice and mice treated with multiple low doses of streptozotocin, as a mouse model of diabetes. INS-1 832/13 beta cells and human islets were treated with IL-1β, IFN-γ and TNF-α to mimic the milieu of early type 1 diabetes. Cells and islets were transfected with miR-21 mimics or inhibitors. Luciferase assays and polyribosomal profiling (PRP) were performed to define miR-21-target interactions. Results Beta cell miR-21 was increased in in vivo models of type 1 diabetes and cytokine-treated cells/islets. miR-21 overexpression decreased cell count and viability, and increased cleaved caspase 3 levels, suggesting increased cell death. In silico prediction tools identified the antiapoptotic mRNA BCL2 as a conserved miR-21 target. Consistent with this, miR-21 overexpression decreased BCL2 transcript and B cell lymphoma 2 (BCL2) protein production, while miR-21 inhibition increased BCL2 protein levels and reduced cleaved caspase 3 levels after cytokine treatment. miR-21-mediated cell death was abrogated in 828/33 cells, which constitutively overexpress Bcl2. Luciferase assays suggested a direct interaction between miR-21 and the BCL2 3' untranslated region. With miR-21 overexpression, PRP revealed a shift of the Bcl2 message towards monosome-associated fractions, indicating inhibition of Bcl2 translation. Finally, overexpression in dispersed human islets confirmed a reduction in BCL2 transcripts and increased cleaved caspase 3 production. Conclusions/interpretation In contrast to the pro-survival role reported in other systems, our results demonstrate that miR-21 increases beta cell death via BCL2 transcript degradation and inhibition of BCL2 translation.

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Cited by

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Authors
Sims, E. K., Lakhter, A. J., Anderson-Baucum, E., Kono, T., Tong, X., & Evans-Molina, C.
Affiliations
Indiana University Indianapolis (US)
Corresponding author
Sims, Emily K.
ORCID
Venue
Diabetologia
Publisher
Springer Science and Business Media LLC
Published
2017-03-09
Volume
60
Issue
6
Pages
1057-1065
ISSN
  • 0012-186X (ISSN-L)
  • 1432-0428
DOI
10.1007/s00125-017-4237-z
PMC
PMC5425307
OpenAlex
W2593075393
Cited by
100
References
1
Citations by year
2026: 8 · 2025: 11 · 2024: 15 · 2023: 5 · 2022: 13 · 2021: 13 · 2020: 10 · 2019: 7 · 2018: 16 · 2017: 2
FWCI
9.81
Type
journal article
Language
en
Open access
Licence
Locations
  • Diabetologia (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • IUScholarWorks (Indiana University) (submittedVersion)
  • Purdue University Indianapolis (Indiana University) (submittedVersion)
  • PMC
Funders
  • National Institute of Diabetes and Digestive and Kidney Diseases (1F32DK104501-01A1, K08DK103983, P30 DK097512, R01 DK093954, T32DK064466)
  • Indiana University Health (IU Health Values Award)
  • National Institutes of Health (UL1TR001108)
  • Veteran’s Administration (Merit Award I01BX001733)
  • Pediatric Endocrine Society Clinical Scholar Award
Topics
  • Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Diabetes and associated disorders
Keywords
miR-21, Bcl-2, beta-cell apoptosis, type 1 diabetes, pancreatic beta cells, 3' untranslated region, caspase-3, inflammatory cytokines, INS-1 832/13 cells, NOD mice, streptozotocin-induced diabetes
MeSH terms
Animals, Diabetes Mellitus, Type 1, Fluorescent Antibody Technique, Humans, Interferon-gamma, Male, Tumor Necrosis Factor-alpha, Immunoblotting, Mice, Inbred NOD, Proto-Oncogene Proteins c-bcl-2, MicroRNAs, Insulin-Secreting Cells, Mice, Rats, Interleukin-1beta

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