Enhancement of L-cystine transport activity and its relation to xCT gene induction at the blood-brain barrier by diethyl maleate treatment

Ken Ichi Hosoya, Masatoshi Tomi, Sumio Ohtsuki, Hitomi Takanaga, Shigeki Saeki, Yoshikatsu Kanai, Hitoshi Endou, Mikihiko Naito, Takashi Tsuruo, Tetsuya Terasaki

研究成果: Article

50 引用 (Scopus)

抄録

The purpose of the present study was to elucidate the mechanism of enhancement of L-cystine uptake at the blood-brain barrier (BBB). The uptake of [ 14C]L-cystine and [ 3H]L-glutamic acid (L-Glu) was determined using a mouse brain endothelial cell line (MBEC4) as an in vitro BBB model. The mRNA levels of L-cystine/L-Glu exchanger, system X c -, which consists of xCT and 4F2hc, were determined by quantitative real-time reverse transcription-polymerase chain reaction analysis. The [ 14C]L-cystine uptake by MBEC4 cells appeared to be mediated via an Na +-independent saturable process. The corresponding Michaelis-Menten constant (K m) was 63.7 μM. In the presence of L-Glu, there was competitive inhibition with an inhibition constant (K i) of 83.5 μM. [ 3H]L-Glu uptake in the absence of Na + was saturable with a K m of 48.1 μM, and it exhibited competitive inhibition with a K i of 24.9 μM in the presence of L-cystine. The mutual inhibition between L-cystine and L-Glu and the type of inhibition suggest that system X c - operates in MBEC4 cells. The xCT and 4F2hc mRNAs were expressed in MBEC4 cells and, following diethyl maleate (DEM) treatment, the xCT mRNA level and L-cystine uptake in MBEC4 cells were enhanced in parallel with an increase in DEM concentration (up to 500 μM). Concomitantly, the glutathione concentration in MBEC4 cells was increased. In conclusion, system X c --mediated L-cystine uptake takes place in MBEC4 cells. L-Cystine transport via system X c - at the BBB is likely to be induced under oxidative stress conditions following DEM treatment due to enhanced transcription of the xCT gene.

元の言語English
ページ(範囲)225-231
ページ数7
ジャーナルJournal of Pharmacology and Experimental Therapeutics
302
発行部数1
DOI
出版物ステータスPublished - 2002
外部発表Yes

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diethyl maleate
Cystine
Blood-Brain Barrier
Genes
Messenger RNA

ASJC Scopus subject areas

  • Pharmacology

これを引用

Enhancement of L-cystine transport activity and its relation to xCT gene induction at the blood-brain barrier by diethyl maleate treatment. / Hosoya, Ken Ichi; Tomi, Masatoshi; Ohtsuki, Sumio; Takanaga, Hitomi; Saeki, Shigeki; Kanai, Yoshikatsu; Endou, Hitoshi; Naito, Mikihiko; Tsuruo, Takashi; Terasaki, Tetsuya.

:: Journal of Pharmacology and Experimental Therapeutics, 巻 302, 番号 1, 2002, p. 225-231.

研究成果: Article

Hosoya, Ken Ichi ; Tomi, Masatoshi ; Ohtsuki, Sumio ; Takanaga, Hitomi ; Saeki, Shigeki ; Kanai, Yoshikatsu ; Endou, Hitoshi ; Naito, Mikihiko ; Tsuruo, Takashi ; Terasaki, Tetsuya. / Enhancement of L-cystine transport activity and its relation to xCT gene induction at the blood-brain barrier by diethyl maleate treatment. :: Journal of Pharmacology and Experimental Therapeutics. 2002 ; 巻 302, 番号 1. pp. 225-231.
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abstract = "The purpose of the present study was to elucidate the mechanism of enhancement of L-cystine uptake at the blood-brain barrier (BBB). The uptake of [ 14C]L-cystine and [ 3H]L-glutamic acid (L-Glu) was determined using a mouse brain endothelial cell line (MBEC4) as an in vitro BBB model. The mRNA levels of L-cystine/L-Glu exchanger, system X c -, which consists of xCT and 4F2hc, were determined by quantitative real-time reverse transcription-polymerase chain reaction analysis. The [ 14C]L-cystine uptake by MBEC4 cells appeared to be mediated via an Na +-independent saturable process. The corresponding Michaelis-Menten constant (K m) was 63.7 μM. In the presence of L-Glu, there was competitive inhibition with an inhibition constant (K i) of 83.5 μM. [ 3H]L-Glu uptake in the absence of Na + was saturable with a K m of 48.1 μM, and it exhibited competitive inhibition with a K i of 24.9 μM in the presence of L-cystine. The mutual inhibition between L-cystine and L-Glu and the type of inhibition suggest that system X c - operates in MBEC4 cells. The xCT and 4F2hc mRNAs were expressed in MBEC4 cells and, following diethyl maleate (DEM) treatment, the xCT mRNA level and L-cystine uptake in MBEC4 cells were enhanced in parallel with an increase in DEM concentration (up to 500 μM). Concomitantly, the glutathione concentration in MBEC4 cells was increased. In conclusion, system X c --mediated L-cystine uptake takes place in MBEC4 cells. L-Cystine transport via system X c - at the BBB is likely to be induced under oxidative stress conditions following DEM treatment due to enhanced transcription of the xCT gene.",
author = "Hosoya, {Ken Ichi} and Masatoshi Tomi and Sumio Ohtsuki and Hitomi Takanaga and Shigeki Saeki and Yoshikatsu Kanai and Hitoshi Endou and Mikihiko Naito and Takashi Tsuruo and Tetsuya Terasaki",
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T1 - Enhancement of L-cystine transport activity and its relation to xCT gene induction at the blood-brain barrier by diethyl maleate treatment

AU - Hosoya, Ken Ichi

AU - Tomi, Masatoshi

AU - Ohtsuki, Sumio

AU - Takanaga, Hitomi

AU - Saeki, Shigeki

AU - Kanai, Yoshikatsu

AU - Endou, Hitoshi

AU - Naito, Mikihiko

AU - Tsuruo, Takashi

AU - Terasaki, Tetsuya

PY - 2002

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N2 - The purpose of the present study was to elucidate the mechanism of enhancement of L-cystine uptake at the blood-brain barrier (BBB). The uptake of [ 14C]L-cystine and [ 3H]L-glutamic acid (L-Glu) was determined using a mouse brain endothelial cell line (MBEC4) as an in vitro BBB model. The mRNA levels of L-cystine/L-Glu exchanger, system X c -, which consists of xCT and 4F2hc, were determined by quantitative real-time reverse transcription-polymerase chain reaction analysis. The [ 14C]L-cystine uptake by MBEC4 cells appeared to be mediated via an Na +-independent saturable process. The corresponding Michaelis-Menten constant (K m) was 63.7 μM. In the presence of L-Glu, there was competitive inhibition with an inhibition constant (K i) of 83.5 μM. [ 3H]L-Glu uptake in the absence of Na + was saturable with a K m of 48.1 μM, and it exhibited competitive inhibition with a K i of 24.9 μM in the presence of L-cystine. The mutual inhibition between L-cystine and L-Glu and the type of inhibition suggest that system X c - operates in MBEC4 cells. The xCT and 4F2hc mRNAs were expressed in MBEC4 cells and, following diethyl maleate (DEM) treatment, the xCT mRNA level and L-cystine uptake in MBEC4 cells were enhanced in parallel with an increase in DEM concentration (up to 500 μM). Concomitantly, the glutathione concentration in MBEC4 cells was increased. In conclusion, system X c --mediated L-cystine uptake takes place in MBEC4 cells. L-Cystine transport via system X c - at the BBB is likely to be induced under oxidative stress conditions following DEM treatment due to enhanced transcription of the xCT gene.

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