Calpain and reactive oxygen species targets Bax for mitochondrial permeabilisation and caspase activation in zerumbone induced apoptosis.
Calpain and reactive oxygen species targets Bax for mitochondrial permeabilisation and caspase activation in zerumbone induced apoptosis.
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DOI：
10.1371/journal.pone.0059350
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
R SK
中科院分区：
文献类型：
--
作者：
Sobhan PK;Seervi M;Deb L;Varghese S;Soman A;Joseph J;Mathew KA;Raghu G;Thomas G;E S;S M;R SK
基于荧光蛋白的信号探针正在成为研究细胞信号传导的有价值的工具，因为它们能够以非侵入性活细胞模式提供时空信息。此前，多种荧光蛋白探针被用来表征不同实验系统中细胞凋亡的关键事件。我们采用基于活细胞图像的方法来可视化癌细胞中由姜黄酮（生姜 Zingiber zerumbet 的活性成分）诱导的细胞凋亡信号传导的关键事件，这使我们能够以时间分辨率分层方式分析显着的细胞凋亡变化。我们的研究证实线粒体透化和细胞色素 c 依赖性 caspase 激活在 zerumbone 诱导的细胞死亡中占主导地位。 Bax 激活是细胞死亡的重要早期事件，由活性氧和钙蛋白酶独立激活。 Zerumbone 未能在 Bax 敲除细胞中诱导细胞凋亡或线粒体透化，并且 Bax 的过度表达增强了 zerumbone 诱导的细胞死亡，证实了 Bax 对于线粒体透化的重要作用。为了防止 Bax 激活和细胞死亡，需要同时抑制活性氧和钙蛋白酶。然而，在 Bcl 2 和 Bcl-XL 过表达细胞中，Zerumbone 诱导的细胞凋亡被阻止，而特异性靶向内质网的 Bcl 2 提供了更多保护。尽管zerumbone治疗下调了XIAP、Survivin和Akt等存活蛋白，但它未能影响PUMA和BIM等促凋亡蛋白。采用多种正常二倍体细胞系来解决Zerumbone的细胞毒活性，一般来说，乳腺上皮细胞、内皮祖细胞和平滑肌细胞对Zerumbone诱导的细胞死亡具有相对抵抗力，并且ROS积累比癌细胞更少。
Fluorescent protein based signaling probes are emerging as valuable tools to study cell signaling because of their ability to provide spatio- temporal information in non invasive live cell mode. Previously, multiple fluorescent protein probes were employed to characterize key events of apoptosis in diverse experimental systems. We have employed a live cell image based approach to visualize the key events of apoptosis signaling induced by zerumbone, the active principle from ginger Zingiber zerumbet, in cancer cells that enabled us to analyze prominent apoptotic changes in a hierarchical manner with temporal resolution. Our studies substantiate that mitochondrial permeabilisation and cytochrome c dependent caspase activation dominate in zerumbone induced cell death. Bax activation, the essential and early event of cell death, is independently activated by reactive oxygen species as well as calpains. Zerumbone failed to induce apoptosis or mitochondrial permeabilisation in Bax knockout cells and over-expression of Bax enhanced cell death induced by zerumbone confirming the essential role of Bax for mitochondrial permeabilsation. Simultaneous inhibition of reactive oxygen species and calpain is required for preventing Bax activation and cell death. However, apoptosis induced by zerumbone was prevented in Bcl 2 and Bcl-XL over-expressing cells, whereas more protection was afforded by Bcl 2 specifically targeted to endoplasmic reticulum. Even though zerumbone treatment down-regulated survival proteins such as XIAP, Survivin and Akt, it failed to affect the pro-apoptotic proteins such as PUMA and BIM. Multiple normal diploid cell lines were employed to address cytotoxic activity of zerumbone and, in general, mammary epithelial cells, endothelial progenitor cells and smooth muscle cells were relatively resistant to zerumbone induced cell death with lesser ROS accumulation than cancer cells.
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影响因子：
5.7
作者：
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通讯作者：
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影响因子：
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影响因子：
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作者：
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