FTY720 stimulated ROS generation and the Sty1/Atf1 signaling pathway in the fission yeast Schizosaccharomyces pombe
FTY720 stimulated ROS generation and the Sty1/Atf1 signaling pathway in the fission yeast Schizosaccharomyces pombe
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DOI：
10.1111/gtc.12134
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发表时间：
2014-04-01
期刊：
影响因子：
2.1
通讯作者：
Sugiura, Reiko
中科院分区：
文献类型：
--
作者：
Hagihara, Kanako;Mizukura, Aya;Sugiura, Reiko
Fingolimod hydrochloride（FTY 720）是第一种免疫调节剂，被称为1-磷酸鞘氨醇（S1 P）受体激动剂。FTY 720还被报道发挥多种生理功能，如抗肿瘤作用、血管生成抑制和Ca 2+动员。在这里，我们表明FTY 720处理诱导活性氧（ROS）积累，并研究FTY 720对应激激活的MAP激酶Spc 1/Sty 1（p38 MAPK的功能同源物）的影响，使用与CRE融合的海肾荧光素酶报告基因构建体，它给出了Atf 1转录活性的精确测量，因此可以作为Spc 1/Sty 1 MAPK信号转导对氧化应激的响应。FTY 720以浓度依赖性方式刺激CRE反应，通过删除Spc 1/Sty 1 MAPK通路的组分，CRE反应显著降低。NAC（N-acetyl-l-cysteine）阻断ROS产生可显著逆转FTY 720诱导的ROS积累、随后的Spc 1/Sty 1 MAPK通路激活和细胞增殖抑制。与野生型细胞相比，缺乏Spc 1/Sty 1 MAPK组分的细胞对FTY 720表现出更高的敏感性，并且在FTY 720处理后表现出更高的ROS水平。因此，我们的研究结果表明，裂变酵母阐明FTY 720介导的信号通路涉及ROS的有用性。
Fingolimod hydrochloride (FTY720) is the first-in-class immune modulator known as sphingosine 1-phosphate (S1P) receptor agonists. FTY720 has also been reported to exert a variety of physiological functions such as antitumor effect, angiogenesis inhibition, and Ca2+ mobilization. Here, we show that FTY720 treatment induced reactive oxygen species (ROS) accumulation, and investigated the effect of FTY720 on the stress-activated MAP kinaseSpc1/Sty1, a functional homologue of p38 MAPK, using a Renilla luciferase reporter construct fused to the CRE, which gives an accurate measure of the transcriptional activity of Atf1 and thus serves as a faithful readout of the Spc1/Sty1 MAPK signaling in response to oxidative stresses. FTY720 stimulated the CRE responses in a concentration-dependent manner, which was markedly reduced by deletion of the components of the Spc1/Sty1 MAPK pathway. The blockade of ROS production by NAC (N-acetyl-l-cysteine) significantly reversed the FTY720-induced ROS accumulation, subsequent activation of the Spc1/Sty1 MAPK pathway, and inhibition of cell proliferation. Cells lacking the components of the Spc1/Sty1 MAPK exhibited higher sensitivity to FTY720 and higher ROS levels upon FTY720 treatment than in wild-type cells. Thus, our results demonstrate the usefulness of fission yeast for elucidating the FTY720-mediated signaling pathways involving ROS.