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Transcriptional Induction of Metallothionein by Tris(pentafluorophenyl)stibane in Cultured Bovine Aortic Endothelial Cells

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ABSTRACT

Vascular endothelial cells cover the luminal surface of blood vessels and contribute to the prevention of vascular disorders such as atherosclerosis. Metallothionein (MT) is a low molecular weight, cysteine-rich, metal-binding, inducible protein, which protects cells from the toxicity of heavy metals and active oxygen species. Endothelial MT is not induced by inorganic zinc. Adequate tools are required to investigate the mechanisms underlying endothelial MT induction. In the present study, we found that an organoantimony compound, tris(pentafluorophenyl)stibane, induces gene expression of MT-1A and MT-2A, which are subisoforms of MT in bovine aortic endothelial cells. The data reveal that MT-1A is induced by activation of both the MTF-1–MRE and Nrf2–ARE pathways, whereas MT-2A expression requires only activation of the MTF-1–MRE pathway. The present data suggest that the original role of MT-1 is to protect cells from heavy metal toxicity and oxidative stress in the biological defense system, while that of MT-2 is to regulate intracellular zinc metabolism.

No MeSH data available.


The Nrf2–ARE pathway mediates P35-induced transcription of endothelial MT-2A. (A,B) ARE-driven transcriptional activity. Vascular endothelial cells transfected with an ARE reporter vector were treated with or without 1, 5, 10, 20, or 30 µM P35 for 12 h, and ARE-driven transcriptional activity was determined by performing the ARE-directed reporter assay. Data are expressed as the mean ± SE of three representative samples, with each sample obtained from three independent experiments. ** p < 0.01 indicates significantly different from the corresponding control; (C) P35-induced transcription of endothelial MT-1A, MT-1E, and MT-2A after Nrf2 knockdown. Vascular endothelial cells transfected with control or Nrf2 siRNA were treated with or without 10 or 30 µM P35 for 12 h. Data are expressed as the mean ± SE of three representative samples, with each sample obtained from three independent experiments. ** p < 0.01 indicates significantly different from the corresponding siControl.
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ijms-17-01381-f008: The Nrf2–ARE pathway mediates P35-induced transcription of endothelial MT-2A. (A,B) ARE-driven transcriptional activity. Vascular endothelial cells transfected with an ARE reporter vector were treated with or without 1, 5, 10, 20, or 30 µM P35 for 12 h, and ARE-driven transcriptional activity was determined by performing the ARE-directed reporter assay. Data are expressed as the mean ± SE of three representative samples, with each sample obtained from three independent experiments. ** p < 0.01 indicates significantly different from the corresponding control; (C) P35-induced transcription of endothelial MT-1A, MT-1E, and MT-2A after Nrf2 knockdown. Vascular endothelial cells transfected with control or Nrf2 siRNA were treated with or without 10 or 30 µM P35 for 12 h. Data are expressed as the mean ± SE of three representative samples, with each sample obtained from three independent experiments. ** p < 0.01 indicates significantly different from the corresponding siControl.

Mentions: Similarly, P35 activated Nrf2 (Figure 8A) and significantly increased ARE-driven transcription in a concentration-dependent manner (Figure 8B). The transcriptional induction of MT-2A by P35 was, however, unaffected in a siRNA-mediated knockdown of Nrf2 (Figure 8B), indicating that the Nrf2–ARE pathway does not transactivate MT-2A induction.


Transcriptional Induction of Metallothionein by Tris(pentafluorophenyl)stibane in Cultured Bovine Aortic Endothelial Cells
The Nrf2–ARE pathway mediates P35-induced transcription of endothelial MT-2A. (A,B) ARE-driven transcriptional activity. Vascular endothelial cells transfected with an ARE reporter vector were treated with or without 1, 5, 10, 20, or 30 µM P35 for 12 h, and ARE-driven transcriptional activity was determined by performing the ARE-directed reporter assay. Data are expressed as the mean ± SE of three representative samples, with each sample obtained from three independent experiments. ** p < 0.01 indicates significantly different from the corresponding control; (C) P35-induced transcription of endothelial MT-1A, MT-1E, and MT-2A after Nrf2 knockdown. Vascular endothelial cells transfected with control or Nrf2 siRNA were treated with or without 10 or 30 µM P35 for 12 h. Data are expressed as the mean ± SE of three representative samples, with each sample obtained from three independent experiments. ** p < 0.01 indicates significantly different from the corresponding siControl.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037661&req=5

ijms-17-01381-f008: The Nrf2–ARE pathway mediates P35-induced transcription of endothelial MT-2A. (A,B) ARE-driven transcriptional activity. Vascular endothelial cells transfected with an ARE reporter vector were treated with or without 1, 5, 10, 20, or 30 µM P35 for 12 h, and ARE-driven transcriptional activity was determined by performing the ARE-directed reporter assay. Data are expressed as the mean ± SE of three representative samples, with each sample obtained from three independent experiments. ** p < 0.01 indicates significantly different from the corresponding control; (C) P35-induced transcription of endothelial MT-1A, MT-1E, and MT-2A after Nrf2 knockdown. Vascular endothelial cells transfected with control or Nrf2 siRNA were treated with or without 10 or 30 µM P35 for 12 h. Data are expressed as the mean ± SE of three representative samples, with each sample obtained from three independent experiments. ** p < 0.01 indicates significantly different from the corresponding siControl.
Mentions: Similarly, P35 activated Nrf2 (Figure 8A) and significantly increased ARE-driven transcription in a concentration-dependent manner (Figure 8B). The transcriptional induction of MT-2A by P35 was, however, unaffected in a siRNA-mediated knockdown of Nrf2 (Figure 8B), indicating that the Nrf2–ARE pathway does not transactivate MT-2A induction.

View Article: PubMed Central - PubMed

ABSTRACT

Vascular endothelial cells cover the luminal surface of blood vessels and contribute to the prevention of vascular disorders such as atherosclerosis. Metallothionein (MT) is a low molecular weight, cysteine-rich, metal-binding, inducible protein, which protects cells from the toxicity of heavy metals and active oxygen species. Endothelial MT is not induced by inorganic zinc. Adequate tools are required to investigate the mechanisms underlying endothelial MT induction. In the present study, we found that an organoantimony compound, tris(pentafluorophenyl)stibane, induces gene expression of MT-1A and MT-2A, which are subisoforms of MT in bovine aortic endothelial cells. The data reveal that MT-1A is induced by activation of both the MTF-1&ndash;MRE and Nrf2&ndash;ARE pathways, whereas MT-2A expression requires only activation of the MTF-1&ndash;MRE pathway. The present data suggest that the original role of MT-1 is to protect cells from heavy metal toxicity and oxidative stress in the biological defense system, while that of MT-2 is to regulate intracellular zinc metabolism.

No MeSH data available.