Withaferin a alters intermediate filament organization, cell shape and behavior.
Withaferin a alters intermediate filament organization, cell shape and behavior.
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DOI：
10.1371/journal.pone.0039065
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Goldman RD
中科院分区：
文献类型：
--
作者：
Grin B;Mahammad S;Wedig T;Cleland MM;Tsai L;Herrmann H;Goldman RD
Withaferin A（WFA）是一种存在于睡茄中的甾体内酯，已在体外显示与中间丝蛋白波形蛋白结合。基于其对波形蛋白的亲和力，有人提出WFA可用作抗肿瘤药物，靶向上调波形蛋白表达的转移性细胞。我们发现，WFA治疗人成纤维细胞迅速重组波形蛋白中间丝（VIF）成核周聚集。这种重组是剂量依赖性的，并伴随着细胞形状的变化，运动性降低和丝氨酸-38处波形蛋白磷酸化的增加。此外，波形蛋白缺乏半胱氨酸-328，建议的WFA结合位点，仍然对WFA敏感，表明该网站是不需要其细胞效应。使用分析超离心，粘度计，电子显微镜和沉降测定，我们表明，WFA在体外对VIF组装没有影响。此外，WFA对波形蛋白不是特异性的，因为它破坏细胞组织并诱导由外周蛋白、神经毒性三联体蛋白和角蛋白组成的几种其他IF网络的核周聚集体。在共表达角蛋白IF和VIF的细胞中，前者对WFA诱导核周聚集体的敏感性显著降低。微管和肌动蛋白/微丝的组织也受到WFA的影响。微管变得波浪形和稀疏和应力纤维的数量似乎增加。在暴露于改变VIF组织和运动性的剂量的WFA 24小时后，细胞经历凋亡。较低剂量的药物不会杀死细胞，但会导致它们衰老。鉴于我们的研究结果，即WFA影响多种IF系统，这些系统在身体的许多组织中表达，因此在将其用作抗癌剂时应谨慎，因为它可能具有使整个生物体衰弱的作用。
Withaferin A (WFA) is a steroidal lactone present in Withania somnifera which has been shown in vitro to bind to the intermediate filament protein, vimentin. Based upon its affinity for vimentin, it has been proposed that WFA can be used as an anti-tumor agent to target metastatic cells which up-regulate vimentin expression. We show that WFA treatment of human fibroblasts rapidly reorganizes vimentin intermediate filaments (VIF) into a perinuclear aggregate. This reorganization is dose dependent and is accompanied by a change in cell shape, decreased motility and an increase in vimentin phosphorylation at serine-38. Furthermore, vimentin lacking cysteine-328, the proposed WFA binding site, remains sensitive to WFA demonstrating that this site is not required for its cellular effects. Using analytical ultracentrifugation, viscometry, electron microscopy and sedimentation assays we show that WFA has no effect on VIF assembly in vitro. Furthermore, WFA is not specific for vimentin as it disrupts the cellular organization and induces perinuclear aggregates of several other IF networks comprised of peripherin, neurofilament-triplet protein, and keratin. In cells co-expressing keratin IF and VIF, the former are significantly less sensitive to WFA with respect to inducing perinuclear aggregates. The organization of microtubules and actin/microfilaments is also affected by WFA. Microtubules become wavier and sparser and the number of stress fibers appears to increase. Following 24 hrs of exposure to doses of WFA that alter VIF organization and motility, cells undergo apoptosis. Lower doses of the drug do not kill cells but cause them to senesce. In light of our findings that WFA affects multiple IF systems, which are expressed in many tissues of the body, caution is warranted in its use as an anti-cancer agent, since it may have debilitating organism-wide effects.
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DOI：
10.1083/jcb.81.3.570
发表时间：
1979-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
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DOI：
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发表时间：
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期刊：
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影响因子：
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影响因子：
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