Modulation of cell surface protein free thiols: a potential novel mechanism of action of the sesquiterpene lactone parthenolide.
Modulation of cell surface protein free thiols: a potential novel mechanism of action of the sesquiterpene lactone parthenolide.
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DOI：
10.1371/journal.pone.0008115
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发表时间：
2009-12-02
期刊：
影响因子：
3.7
通讯作者：
Bernstein SH
中科院分区：
文献类型：
--
作者：
Skalska J;Brookes PS;Nadtochiy SM;Hilchey SP;Jordan CT;Guzman ML;Maggirwar SB;Briehl MM;Bernstein SH
靶向细胞内氧化还原通路作为癌症的一种治疗方法一直受到人们的极大兴趣。鉴于最近的数据表明细胞外蛋白硫醇基团(即表面硫醇)的氧化还原状态影响细胞行为，我们假设氧化还原活性抗癌药物将调节表面蛋白硫醇。为了验证这一假设，我们使用了倍半萜内酯巴特内酯，一种已知的抗癌药物。用Alexa Fluor 633 C5马来酰亚胺染料和N-(生物素)-N-(碘乙酰基)乙二胺(BIAM)分别用流式细胞术和免疫印迹法标记游离硫醇，结果表明，小白菊内酯能降低格兰特外套淋巴瘤细胞表面游离硫醇的水平。此外，我们使用免疫沉淀技术来鉴定中心氧化还原调节剂硫氧还蛋白，它是巴特内酯修饰的表面蛋白硫醇靶标之一。为了研究Parthenolide诱导的表面蛋白硫醇修饰的功能作用，我们在Parthenolide暴露之前用细胞不透性谷胱甘肽预先处理Granta细胞，结果表明：(A)抑制Parthenolide与表面硫醇的相互作用；(B)抑制Parthenolide介导的JNK的激活和抑制NFκB，这两个成熟的机制是Parthenolide的活性和；(C)阻断Parthenolide的细胞毒活性。GSH对菊内酯诱导的细胞内氧自由基的产生没有影响，这支持了GSH对细胞内氧化还原没有影响的事实。综上所述，这些数据支持了这样一种可能性，即谷胱甘肽可能通过直接抑制巴豆内酯对表面硫醇的调节而抑制其对jnk、nfκB和细胞死亡的影响。基于这些数据，我们推测，菊内酯抗淋巴瘤活性的一个成分来自于它改变关键表面硫醇的氧化还原状态的能力。此外，我们认为癌细胞表面硫醇可能是重要的和新的治疗靶点。
There has been much interest in targeting intracellular redox pathways as a therapeutic approach for cancer. Given recent data to suggest that the redox status of extracellular protein thiol groups (i.e. exofacial thiols) effects cell behavior, we hypothesized that redox active anti-cancer agents would modulate exofacial protein thiols. To test this hypothesis, we used the sesquiterpene lactone parthenolide, a known anti-cancer agent. Using flow cytometry, and western blotting to label free thiols with Alexa Fluor 633 C5 maleimide dye and N-(biotinoyl)-N-(iodoacetyl) ethylendiamine (BIAM), respectively, we show that parthenolide decreases the level of free exofacial thiols on Granta mantle lymphoma cells. In addition, we used immuno-precipitation techniques to identify the central redox regulator thioredoxin, as one of the surface protein thiol targets modified by parthenolide. To examine the functional role of parthenolide induced surface protein thiol modification, we pretreated Granta cells with cell impermeable glutathione (GSH), prior to exposure to parthenolide, and showed that GSH pretreatment; (a) inhibited the interaction of parthenolide with exofacial thiols; (b) inhibited parthenolide mediated activation of JNK and inhibition of NFκB, two well established mechanisms of parthenolide activity and; (c) blocked the cytotoxic activity of parthenolide. That GSH had no effect on the parthenolide induced generation of intracellular reactive oxygen species supports the fact that GSH had no effect on intracellular redox. Together these data support the likelihood that GSH inhibits the effect of parthenolide on JNK, NFκB and cell death through its direct inhibition of parthenolide's modulation of exofacial thiols. Based on these data, we postulate that one component of parthenolide's anti-lymphoma activity derives from its ability to modify the redox state of critical exofacial thiols. Further, we propose that cancer cell exofacial thiols may be important and novel targets for therapy.
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影响因子：
11.4
作者：
Jadayel, DM;Lukas, J;Catovsky, D
通讯作者：
Catovsky, D
影响因子：
2.6
作者：
Ray, Denise M.;Morse, Kimberly M.;Bernstein, Steven H.
通讯作者：
Bernstein, Steven H.
影响因子：
20.3
作者：
Guzman, ML;Rossi, RM;Jordan, CT
通讯作者：
Jordan, CT
DOI：
10.1073/pnas.2434516100
发表时间：
2003-12-09
影响因子：
11.1
作者：
Laragione, T;Bonetto, V;Ghezzi, P
通讯作者：
Ghezzi, P
影响因子：
11.2
作者：
Brookes, Paul S.;Morse, Kimberly;Bernstein, Steven H.
通讯作者：
Bernstein, Steven H.