Tributyltin induces G2/M cell cycle arrest via NAD+-dependent isocitrate dehydrogenase in human embryonic carcinoma cells

Miki Asanagi, Shigeru Yamada, Naoya Hirata, Hiroshi Itagaki, Yaichiro Kotake, Yuko Sekino, Yasunari Kanda

  • 2 Citations

Abstract

Organotin compounds, such as tributyltin (TBT), are well-known endocrine-disrupting chemicals (EDCs). We have recently reported that TBT induces growth arrest in the human embryonic carcinoma cell line NT2/D1 at nanomolar levels by inhibiting NAD+-dependent isocitrate dehydrogenase (NAD-IDH), which catalyzes the irreversible conversion of isocitrate to α-ketoglutarate. However, the molecular mechanisms by which NAD-IDH mediates TBT toxicity remain unclear. In the present study, we examined whether TBT at nanomolar levels affects cell cycle progression in NT2/D1 cells. Propidium iodide staining revealed that TBT reduced the ratio of cells in the G1 phase and increased the ratio of cells in the G2/M phase. TBT also reduced cell division cycle 25C (cdc25C) and cyclin B1, which are key regulators of G2/M progression. Furthermore, apigenin, an inhibitor of NAD-IDH, mimicked the effects of TBT. The G2/M arrest induced by TBT was abolished by NAD-IDHα knockdown. Treatment with a cellpermeable α-ketoglutarate analogue recovered the effect of TBT, suggesting the involvement of NADIDH. Taken together, our data suggest that TBT at nanomolar levels induced G2/M cell cycle arrest via NAD-IDH in NT2/D1 cells. Thus, cell cycle analysis in embryonic cells could be used to assess cytotoxicity associated with nanomolar level exposure of EDCs.

Original languageEnglish
Pages (from-to)207-215
Number of pages9
JournalJournal of Toxicological Sciences
Volume41
Issue number2
DOIs
StatePublished - 2016 Mar 10

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G2 Phase Cell Cycle Checkpoints
Isocitrate Dehydrogenase
NAD
Carcinoma
Cell Cycle
Endocrine Disruptors
Organotin Compounds
Cyclin B1
Apigenin
Propidium
G2 Phase
G1 Phase
Cell Division
Staining and Labeling
Cell Line

Keywords

  • Cell cycle
  • Embryonic carcinoma cells
  • Isocitrate dehydrogenase
  • Tributyltin

ASJC Scopus subject areas

  • Toxicology

Cite this

Tributyltin induces G2/M cell cycle arrest via NAD+-dependent isocitrate dehydrogenase in human embryonic carcinoma cells. / Asanagi, Miki; Yamada, Shigeru; Hirata, Naoya; Itagaki, Hiroshi; Kotake, Yaichiro; Sekino, Yuko; Kanda, Yasunari.

In: Journal of Toxicological Sciences, Vol. 41, No. 2, 10.03.2016, p. 207-215.

Research output: Contribution to journalArticle

Asanagi, Miki; Yamada, Shigeru; Hirata, Naoya; Itagaki, Hiroshi; Kotake, Yaichiro; Sekino, Yuko; Kanda, Yasunari / Tributyltin induces G2/M cell cycle arrest via NAD+-dependent isocitrate dehydrogenase in human embryonic carcinoma cells.

In: Journal of Toxicological Sciences, Vol. 41, No. 2, 10.03.2016, p. 207-215.

Research output: Contribution to journalArticle

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author = "Miki Asanagi and Shigeru Yamada and Naoya Hirata and Hiroshi Itagaki and Yaichiro Kotake and Yuko Sekino and Yasunari Kanda",
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AU - Asanagi,Miki

AU - Yamada,Shigeru

AU - Hirata,Naoya

AU - Itagaki,Hiroshi

AU - Kotake,Yaichiro

AU - Sekino,Yuko

AU - Kanda,Yasunari

PY - 2016/3/10

Y1 - 2016/3/10

N2 - Organotin compounds, such as tributyltin (TBT), are well-known endocrine-disrupting chemicals (EDCs). We have recently reported that TBT induces growth arrest in the human embryonic carcinoma cell line NT2/D1 at nanomolar levels by inhibiting NAD+-dependent isocitrate dehydrogenase (NAD-IDH), which catalyzes the irreversible conversion of isocitrate to α-ketoglutarate. However, the molecular mechanisms by which NAD-IDH mediates TBT toxicity remain unclear. In the present study, we examined whether TBT at nanomolar levels affects cell cycle progression in NT2/D1 cells. Propidium iodide staining revealed that TBT reduced the ratio of cells in the G1 phase and increased the ratio of cells in the G2/M phase. TBT also reduced cell division cycle 25C (cdc25C) and cyclin B1, which are key regulators of G2/M progression. Furthermore, apigenin, an inhibitor of NAD-IDH, mimicked the effects of TBT. The G2/M arrest induced by TBT was abolished by NAD-IDHα knockdown. Treatment with a cellpermeable α-ketoglutarate analogue recovered the effect of TBT, suggesting the involvement of NADIDH. Taken together, our data suggest that TBT at nanomolar levels induced G2/M cell cycle arrest via NAD-IDH in NT2/D1 cells. Thus, cell cycle analysis in embryonic cells could be used to assess cytotoxicity associated with nanomolar level exposure of EDCs.

AB - Organotin compounds, such as tributyltin (TBT), are well-known endocrine-disrupting chemicals (EDCs). We have recently reported that TBT induces growth arrest in the human embryonic carcinoma cell line NT2/D1 at nanomolar levels by inhibiting NAD+-dependent isocitrate dehydrogenase (NAD-IDH), which catalyzes the irreversible conversion of isocitrate to α-ketoglutarate. However, the molecular mechanisms by which NAD-IDH mediates TBT toxicity remain unclear. In the present study, we examined whether TBT at nanomolar levels affects cell cycle progression in NT2/D1 cells. Propidium iodide staining revealed that TBT reduced the ratio of cells in the G1 phase and increased the ratio of cells in the G2/M phase. TBT also reduced cell division cycle 25C (cdc25C) and cyclin B1, which are key regulators of G2/M progression. Furthermore, apigenin, an inhibitor of NAD-IDH, mimicked the effects of TBT. The G2/M arrest induced by TBT was abolished by NAD-IDHα knockdown. Treatment with a cellpermeable α-ketoglutarate analogue recovered the effect of TBT, suggesting the involvement of NADIDH. Taken together, our data suggest that TBT at nanomolar levels induced G2/M cell cycle arrest via NAD-IDH in NT2/D1 cells. Thus, cell cycle analysis in embryonic cells could be used to assess cytotoxicity associated with nanomolar level exposure of EDCs.

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