Long-term pancreatic beta cell exposure to high levels of glucose but not palmitate induces DNA methylation within the insulin gene promoter and represses transcriptional activity

Kota Ishikawa, Shin Tsunekawa, Makoto Ikeniwa, Takako Izumoto, Atsushi Iida, Hidetada Ogata, Eita Uenishi, Yusuke Seino, Nobuaki Ozaki, Yoshihisa Sugimura, Yoji Hamada, Akio Kuroda, Keiko Shinjo, Yutaka Kondo, Yutaka Oiso

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18 Citations (Scopus)

Abstract

Recent studies have implicated epigenetics in the pathophysiology of diabetes. Furthermore, DNA methylation, which irreversibly deactivates gene transcription, of the insulin promoter, particularly the cAMP response element, is increased in diabetes patients. However, the underlying mechanism remains unclear. We aimed to investigate insulin promoter DNA methylation in an over-nutrition state. INS-1 cells, the rat pancreatic beta cell line, were cultured under normal-culture-glucose (11.2mmol/l) or experimental-high-glucose (22.4mmol/l) conditions for 14 days, with or without 0.4 mmol/l palmitate. DNA methylation of the rat insulin 1 gene (Ins1) promoter was investigated using bisulfite sequencing and pyrosequencing analysis. Experimental-high-glucose conditions significantly suppressed insulin mRNA and increased DNA methylation at all five CpG sites within the Ins1 promoter, including the cAMP response element, in a time-dependent and glucose concentration-dependent manner. DNA methylation under experimental-high-glucose conditions was unique to the Ins1 promoter; however, palmitate did not affect DNA methylation. Artificial methylation of Ins1 promoter significantly suppressed promoter-driven luciferase activity, and a DNA methylation inhibitor significantly improved insulin mRNA suppression by experimental-high-glucose conditions. Experimental-high-glucose conditions significantly increased DNA methyltransferase activity and decreased ten-eleven-translocation methylcytosine dioxygenase activity. Oxidative stress and endoplasmic reticulumstress did not affect DNA methylation of the Ins1 promoter. High glucose but not palmitate increased ectopic triacylglycerol accumulation parallel to DNA methylation. Metformin upregulated insulin gene expression and suppressed DNA methylation and ectopic triacylglycerol accumulation. Finally, DNA methylation of the Ins1 promoter increased in isolated islets from Zucker diabetic fatty rats. This study helps to clarify the effect of an over-nutrition state on DNA methylation of the Ins1 promoter in pancreatic beta cells. It provides new insights into the irreversible pathophysiology of diabetes.

Original languageEnglish
Article numbere0115350
JournalPloS one
Volume10
Issue number2
DOIs
Publication statusPublished - 06-02-2015

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Palmitates
palmitates
islets of Langerhans
Insulin-Secreting Cells
DNA methylation
DNA Methylation
insulin
Genes
promoter regions
Insulin
Glucose
glucose
genes
Medical problems
diabetes
Rats
response elements
Response Elements
Nutrition
pathophysiology

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Ishikawa, Kota ; Tsunekawa, Shin ; Ikeniwa, Makoto ; Izumoto, Takako ; Iida, Atsushi ; Ogata, Hidetada ; Uenishi, Eita ; Seino, Yusuke ; Ozaki, Nobuaki ; Sugimura, Yoshihisa ; Hamada, Yoji ; Kuroda, Akio ; Shinjo, Keiko ; Kondo, Yutaka ; Oiso, Yutaka. / Long-term pancreatic beta cell exposure to high levels of glucose but not palmitate induces DNA methylation within the insulin gene promoter and represses transcriptional activity. In: PloS one. 2015 ; Vol. 10, No. 2.
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abstract = "Recent studies have implicated epigenetics in the pathophysiology of diabetes. Furthermore, DNA methylation, which irreversibly deactivates gene transcription, of the insulin promoter, particularly the cAMP response element, is increased in diabetes patients. However, the underlying mechanism remains unclear. We aimed to investigate insulin promoter DNA methylation in an over-nutrition state. INS-1 cells, the rat pancreatic beta cell line, were cultured under normal-culture-glucose (11.2mmol/l) or experimental-high-glucose (22.4mmol/l) conditions for 14 days, with or without 0.4 mmol/l palmitate. DNA methylation of the rat insulin 1 gene (Ins1) promoter was investigated using bisulfite sequencing and pyrosequencing analysis. Experimental-high-glucose conditions significantly suppressed insulin mRNA and increased DNA methylation at all five CpG sites within the Ins1 promoter, including the cAMP response element, in a time-dependent and glucose concentration-dependent manner. DNA methylation under experimental-high-glucose conditions was unique to the Ins1 promoter; however, palmitate did not affect DNA methylation. Artificial methylation of Ins1 promoter significantly suppressed promoter-driven luciferase activity, and a DNA methylation inhibitor significantly improved insulin mRNA suppression by experimental-high-glucose conditions. Experimental-high-glucose conditions significantly increased DNA methyltransferase activity and decreased ten-eleven-translocation methylcytosine dioxygenase activity. Oxidative stress and endoplasmic reticulumstress did not affect DNA methylation of the Ins1 promoter. High glucose but not palmitate increased ectopic triacylglycerol accumulation parallel to DNA methylation. Metformin upregulated insulin gene expression and suppressed DNA methylation and ectopic triacylglycerol accumulation. Finally, DNA methylation of the Ins1 promoter increased in isolated islets from Zucker diabetic fatty rats. This study helps to clarify the effect of an over-nutrition state on DNA methylation of the Ins1 promoter in pancreatic beta cells. It provides new insights into the irreversible pathophysiology of diabetes.",
author = "Kota Ishikawa and Shin Tsunekawa and Makoto Ikeniwa and Takako Izumoto and Atsushi Iida and Hidetada Ogata and Eita Uenishi and Yusuke Seino and Nobuaki Ozaki and Yoshihisa Sugimura and Yoji Hamada and Akio Kuroda and Keiko Shinjo and Yutaka Kondo and Yutaka Oiso",
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Ishikawa, K, Tsunekawa, S, Ikeniwa, M, Izumoto, T, Iida, A, Ogata, H, Uenishi, E, Seino, Y, Ozaki, N, Sugimura, Y, Hamada, Y, Kuroda, A, Shinjo, K, Kondo, Y & Oiso, Y 2015, 'Long-term pancreatic beta cell exposure to high levels of glucose but not palmitate induces DNA methylation within the insulin gene promoter and represses transcriptional activity', PloS one, vol. 10, no. 2, e0115350. https://doi.org/10.1371/journal.pone.0115350

Long-term pancreatic beta cell exposure to high levels of glucose but not palmitate induces DNA methylation within the insulin gene promoter and represses transcriptional activity. / Ishikawa, Kota; Tsunekawa, Shin; Ikeniwa, Makoto; Izumoto, Takako; Iida, Atsushi; Ogata, Hidetada; Uenishi, Eita; Seino, Yusuke; Ozaki, Nobuaki; Sugimura, Yoshihisa; Hamada, Yoji; Kuroda, Akio; Shinjo, Keiko; Kondo, Yutaka; Oiso, Yutaka.

In: PloS one, Vol. 10, No. 2, e0115350, 06.02.2015.

Research output: Contribution to journalArticle

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T1 - Long-term pancreatic beta cell exposure to high levels of glucose but not palmitate induces DNA methylation within the insulin gene promoter and represses transcriptional activity

AU - Ishikawa, Kota

AU - Tsunekawa, Shin

AU - Ikeniwa, Makoto

AU - Izumoto, Takako

AU - Iida, Atsushi

AU - Ogata, Hidetada

AU - Uenishi, Eita

AU - Seino, Yusuke

AU - Ozaki, Nobuaki

AU - Sugimura, Yoshihisa

AU - Hamada, Yoji

AU - Kuroda, Akio

AU - Shinjo, Keiko

AU - Kondo, Yutaka

AU - Oiso, Yutaka

PY - 2015/2/6

Y1 - 2015/2/6

N2 - Recent studies have implicated epigenetics in the pathophysiology of diabetes. Furthermore, DNA methylation, which irreversibly deactivates gene transcription, of the insulin promoter, particularly the cAMP response element, is increased in diabetes patients. However, the underlying mechanism remains unclear. We aimed to investigate insulin promoter DNA methylation in an over-nutrition state. INS-1 cells, the rat pancreatic beta cell line, were cultured under normal-culture-glucose (11.2mmol/l) or experimental-high-glucose (22.4mmol/l) conditions for 14 days, with or without 0.4 mmol/l palmitate. DNA methylation of the rat insulin 1 gene (Ins1) promoter was investigated using bisulfite sequencing and pyrosequencing analysis. Experimental-high-glucose conditions significantly suppressed insulin mRNA and increased DNA methylation at all five CpG sites within the Ins1 promoter, including the cAMP response element, in a time-dependent and glucose concentration-dependent manner. DNA methylation under experimental-high-glucose conditions was unique to the Ins1 promoter; however, palmitate did not affect DNA methylation. Artificial methylation of Ins1 promoter significantly suppressed promoter-driven luciferase activity, and a DNA methylation inhibitor significantly improved insulin mRNA suppression by experimental-high-glucose conditions. Experimental-high-glucose conditions significantly increased DNA methyltransferase activity and decreased ten-eleven-translocation methylcytosine dioxygenase activity. Oxidative stress and endoplasmic reticulumstress did not affect DNA methylation of the Ins1 promoter. High glucose but not palmitate increased ectopic triacylglycerol accumulation parallel to DNA methylation. Metformin upregulated insulin gene expression and suppressed DNA methylation and ectopic triacylglycerol accumulation. Finally, DNA methylation of the Ins1 promoter increased in isolated islets from Zucker diabetic fatty rats. This study helps to clarify the effect of an over-nutrition state on DNA methylation of the Ins1 promoter in pancreatic beta cells. It provides new insights into the irreversible pathophysiology of diabetes.

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