Imaging in real-time with FRET the redox response of tumorigenic cells to glutathione perturbations in a microscale flow.
Imaging in real-time with FRET the redox response of tumorigenic cells to glutathione perturbations in a microscale flow.
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DOI：
10.1039/c0ib00071j
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发表时间：
2011-03
期刊：
影响因子：
--
通讯作者：
Timp G
中科院分区：
文献类型：
--
作者：
Lin C;Kolossov VL;Tsvid G;Trump L;Henry JJ;Henderson JL;Rund LA;Kenis PJ;Schook LB;Gaskins HR;Timp G
尽管选择性氧化还原调节策略对癌症治疗有潜在的好处，但由于很难随着时间的推移测量细胞内的氧化还原电位，测试治疗的有效方法仍然难以实现。在这份报告中，我们加入了一种新的基于FRET的生物传感器，可以实时跟踪转化为致瘤细胞并在微流控通道中培养的氧化还原敏感过程。利用微流控网络控制细胞附近的微尺度流动，同时外源输送药物。随后，测试了氧化还原稳态电路，即还原型谷胱甘肽(GSH)/氧化谷胱甘肽(GSSG)对硫醇氧化剂联胺和两种用于癌症治疗的药物：BSO(L-丁硫氨酸-[SR]-亚磺胺)和BCNU(卡莫司汀)的响应。这些实验的主要结果是实时比较单个活细胞中GSH的时间耗竭和恢复情况。这些数据表明，哺乳动物细胞能够在急性氧化损伤消除后的几分钟内恢复减少的细胞内氧化还原环境。(I)BSO抑制GSH的生物合成；(Ii)BCNU使谷胱甘肽还原酶失活；(Iii)在致瘤细胞中，相对于同基因的非致瘤对照细胞系，这种恢复明显延迟。
Despite the potential benefits of selective redox-modulating strategies for cancer therapy, an efficacious methodology for testing therapies remains elusive because of the difficulty in measuring intracellular redox potentials over time. In this report, we have incorporated a new FRET-based biosensor to follow in real time redox-sensitive processes in cells transformed to be tumorigenic and cultured in a microfluidic channel. A microfluidic network was used to control micro-scale flow near the cells and at the same time deliver drugs exogenously. Subsequently, the response of a redox homeostasis circuit was tested, namely reduced glutathione (GSH)/oxidized glutathione(GSSG), to diamide, a thiol oxidant, and two drugs used for cancer therapies: BSO (l-buthionine-[SR]-sulfoximine) and BCNU (carmustine). The main outcome from these experiments is a comparison of the temporal depletion and recovery of GSH in single living cells in real-time. These data demonstrate that mammalian cells are capable of restoring a reduced intracellular redox environment in minutes after an acute oxidative insult is removed. This recovery is significantly delayed by (i) the inhibition of GSH biosynthesis by BSO; (ii) the inactivation of glutathione reductase by BCNU; and (iii) in tumorigenic cells relative to an isogenic non-tumorigenic control cell line.
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影响因子：
6.1
作者：
Adler M;Polinkovsky M;Gutierrez E;Groisman A
通讯作者：
Groisman A
影响因子：
8
作者：
Adam, S. J.;Rund, L. A.;Counter, C. M.
通讯作者：
Counter, C. M.
影响因子：
2.9
作者：
Kamencic, H;Lyon, A;Juurlink, BHJ
通讯作者：
Juurlink, BHJ
影响因子：
48
作者：
Gutscher, Marcus;Pauleau, Anne-Laure;Dick, Tobias P.
通讯作者：
Dick, Tobias P.
影响因子：
4.6
作者：
Conour, JE;Graham, WV;Gaskins, HR
通讯作者：
Gaskins, HR