Iron suppresses ovarian granulosa cell proliferation and arrests cell cycle through regulating p38 mitogen-activated protein kinase/p53/p21 pathway

Mei Jou Chen, Chia Hong Chou, Chia Tung Shun, Tsui Lien Mao, Wen Fen Wen, Chin Der Chen, Shee Uan Chen, Yu Shih Yang, Hong Nerng Ho

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Iron is an essential nutrient that may exert toxic effects when it accumulates in tissues. Little is known regarding its effects on gonadal function. Both Fe 2+ and Fe 3+ could be released from iron deposition. We employed mouse nonluteinized granulosa cell for in vitro studies and human ovarian tissues for Prussian blue and immunohistochemical staining to identify the iron deposition and effect in vivo. After treatment with FeSO 4 -7H 2 O or FeCl 3 in granulosa cell cultured with folliclestimulating hormone (FSH) for 48 h, we found that Fe 2+ significantly suppressed FSH-induced granulosa cell proliferation and arrested the cell cycle at the G2/M phase by cell proliferation assay and flow cytometry. Fe 2+ significantly increased intracellular reactive oxygen species (ROS) and ferritin levels of mouse granulosa cells. The increases in p21 and p53 messenger RNA and protein expression facilitated by Fe 2+ treatment in mouse granulosa cells were significantly suppressed by separate treatments with p53 small interfering RNA and p38 mitogen-activated protein kinase (MAPK) inhibitors. An ROS inhibitor downregulated Fe 2+ -induced increases in p38MAPK expression in mouse granulosa cells. Quantitative analysis of immunohistochemical staining revealed that human ovarian tissue sections with positive Prussian blue staining had lower levels of proliferating cell nuclear antigen expression, but higher levels of p21, p53, and CDC25C expression than those with negative Prussian blue staining. Conclusively, Fe 2+ could directly arrest the cell cycle and inhibit granulosa cell proliferation by regulating the ROS-mediated p38MAPK/p53/p21 pathway. Therefore, iron can directly affect female gonadal function.

Original languageEnglish
Pages (from-to)438-448
Number of pages11
JournalBiology of Reproduction
Volume97
Issue number3
DOIs
Publication statusPublished - Jan 1 2017
Externally publishedYes

Fingerprint

Granulosa Cells
p38 Mitogen-Activated Protein Kinases
Cell Cycle Checkpoints
Iron
Cell Proliferation
Staining and Labeling
Reactive Oxygen Species
Hormones
G2 Phase
Poisons
Proliferating Cell Nuclear Antigen
Protein Kinase Inhibitors
Ferritins
Cell Division
Small Interfering RNA
Cell Cycle
Flow Cytometry
Down-Regulation
Food
Messenger RNA

Keywords

  • Cell cycle
  • Granulosa cells
  • Iron
  • Ovary
  • Reactive oxygen specie

ASJC Scopus subject areas

  • Reproductive Medicine
  • Cell Biology

Cite this

Iron suppresses ovarian granulosa cell proliferation and arrests cell cycle through regulating p38 mitogen-activated protein kinase/p53/p21 pathway. / Chen, Mei Jou; Chou, Chia Hong; Shun, Chia Tung; Mao, Tsui Lien; Wen, Wen Fen; Chen, Chin Der; Chen, Shee Uan; Yang, Yu Shih; Ho, Hong Nerng.

In: Biology of Reproduction, Vol. 97, No. 3, 01.01.2017, p. 438-448.

Research output: Contribution to journalArticle

Chen, Mei Jou ; Chou, Chia Hong ; Shun, Chia Tung ; Mao, Tsui Lien ; Wen, Wen Fen ; Chen, Chin Der ; Chen, Shee Uan ; Yang, Yu Shih ; Ho, Hong Nerng. / Iron suppresses ovarian granulosa cell proliferation and arrests cell cycle through regulating p38 mitogen-activated protein kinase/p53/p21 pathway. In: Biology of Reproduction. 2017 ; Vol. 97, No. 3. pp. 438-448.
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AU - Chou, Chia Hong

AU - Shun, Chia Tung

AU - Mao, Tsui Lien

AU - Wen, Wen Fen

AU - Chen, Chin Der

AU - Chen, Shee Uan

AU - Yang, Yu Shih

AU - Ho, Hong Nerng

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N2 - Iron is an essential nutrient that may exert toxic effects when it accumulates in tissues. Little is known regarding its effects on gonadal function. Both Fe 2+ and Fe 3+ could be released from iron deposition. We employed mouse nonluteinized granulosa cell for in vitro studies and human ovarian tissues for Prussian blue and immunohistochemical staining to identify the iron deposition and effect in vivo. After treatment with FeSO 4 -7H 2 O or FeCl 3 in granulosa cell cultured with folliclestimulating hormone (FSH) for 48 h, we found that Fe 2+ significantly suppressed FSH-induced granulosa cell proliferation and arrested the cell cycle at the G2/M phase by cell proliferation assay and flow cytometry. Fe 2+ significantly increased intracellular reactive oxygen species (ROS) and ferritin levels of mouse granulosa cells. The increases in p21 and p53 messenger RNA and protein expression facilitated by Fe 2+ treatment in mouse granulosa cells were significantly suppressed by separate treatments with p53 small interfering RNA and p38 mitogen-activated protein kinase (MAPK) inhibitors. An ROS inhibitor downregulated Fe 2+ -induced increases in p38MAPK expression in mouse granulosa cells. Quantitative analysis of immunohistochemical staining revealed that human ovarian tissue sections with positive Prussian blue staining had lower levels of proliferating cell nuclear antigen expression, but higher levels of p21, p53, and CDC25C expression than those with negative Prussian blue staining. Conclusively, Fe 2+ could directly arrest the cell cycle and inhibit granulosa cell proliferation by regulating the ROS-mediated p38MAPK/p53/p21 pathway. Therefore, iron can directly affect female gonadal function.

AB - Iron is an essential nutrient that may exert toxic effects when it accumulates in tissues. Little is known regarding its effects on gonadal function. Both Fe 2+ and Fe 3+ could be released from iron deposition. We employed mouse nonluteinized granulosa cell for in vitro studies and human ovarian tissues for Prussian blue and immunohistochemical staining to identify the iron deposition and effect in vivo. After treatment with FeSO 4 -7H 2 O or FeCl 3 in granulosa cell cultured with folliclestimulating hormone (FSH) for 48 h, we found that Fe 2+ significantly suppressed FSH-induced granulosa cell proliferation and arrested the cell cycle at the G2/M phase by cell proliferation assay and flow cytometry. Fe 2+ significantly increased intracellular reactive oxygen species (ROS) and ferritin levels of mouse granulosa cells. The increases in p21 and p53 messenger RNA and protein expression facilitated by Fe 2+ treatment in mouse granulosa cells were significantly suppressed by separate treatments with p53 small interfering RNA and p38 mitogen-activated protein kinase (MAPK) inhibitors. An ROS inhibitor downregulated Fe 2+ -induced increases in p38MAPK expression in mouse granulosa cells. Quantitative analysis of immunohistochemical staining revealed that human ovarian tissue sections with positive Prussian blue staining had lower levels of proliferating cell nuclear antigen expression, but higher levels of p21, p53, and CDC25C expression than those with negative Prussian blue staining. Conclusively, Fe 2+ could directly arrest the cell cycle and inhibit granulosa cell proliferation by regulating the ROS-mediated p38MAPK/p53/p21 pathway. Therefore, iron can directly affect female gonadal function.

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KW - Ovary

KW - Reactive oxygen specie

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