Withaferin A Induces Cell Death Selectively in Androgen-Independent Prostate Cancer Cells but Not in Normal Fibroblast Cells.
Withaferin A Induces Cell Death Selectively in Androgen-Independent Prostate Cancer Cells but Not in Normal Fibroblast Cells.
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DOI：
10.1371/journal.pone.0134137
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Nojima H
中科院分区：
文献类型：
--
作者：
Nishikawa Y;Okuzaki D;Fukushima K;Mukai S;Ohno S;Ozaki Y;Yabuta N;Nojima H
Withaferin A(Withaferin A，Withaferin A)是印度草本植物睡莲的主要生物活性成分，可诱导多种肿瘤细胞死亡(凋亡/坏死)，但其细胞毒性的分子机制尚不清楚。我们报道了2μM WA选择性地诱导雄激素不敏感的PC-3和DU-14 5前列腺癌细胞死亡，而对雄激素敏感的LNCaP前列腺癌细胞和正常人成纤维细胞(TIG-1和KD)的毒性较小。在椭球形成液中对PC-3细胞也有明显的杀伤作用。DNA芯片分析显示，Wa显著增加PC-3和DU-145细胞c-Fos和11个热休克蛋白(HSPs)的mRNA水平，但对LNCaP和TIG-1细胞无明显影响。Western分析显示，c-Fos表达增加，抗凋亡蛋白c-FLIP表达降低(L)。HSPA6和HSP70等HSPs的表达显著增加；然而，由于siRNA介导的HSF-1(HSP诱导的转录因子)的缺失降低了PC-3细胞的存活率，这些热休克基因可能参与了保护细胞死亡的过程。此外，WA可诱导PC-3和DU-145细胞产生ROS，但不能诱导正常成纤维细胞产生ROS。免疫细胞化学和免疫电子显微镜显示，WA破坏了Vimentin细胞骨架，可能诱导了ROS的产生、c-Fos的表达和c-flip(L)的抑制。这些观察结果表明，随后发生的多个事件以及波形蛋白细胞骨架的破坏在WA介导的细胞死亡中起着关键作用。
Withaferin A (WA), a major bioactive component of the Indian herb Withania somnifera, induces cell death (apoptosis/necrosis) in multiple types of tumor cells, but the molecular mechanism underlying this cytotoxicity remains elusive. We report here that 2 μM WA induced cell death selectively in androgen-insensitive PC-3 and DU-145 prostate adenocarcinoma cells, whereas its toxicity was less severe in androgen-sensitive LNCaP prostate adenocarcinoma cells and normal human fibroblasts (TIG-1 and KD). WA also killed PC-3 cells in spheroid-forming medium. DNA microarray analysis revealed that WA significantly increased mRNA levels of c-Fos and 11 heat-shock proteins (HSPs) in PC-3 and DU-145, but not in LNCaP and TIG-1. Western analysis revealed increased expression of c-Fos and reduced expression of the anti-apoptotic protein c-FLIP(L). Expression of HSPs such as HSPA6 and Hsp70 was conspicuously elevated; however, because siRNA-mediated depletion of HSF-1, an HSP-inducing transcription factor, reduced PC-3 cell viability, it is likely that these heat-shock genes were involved in protecting against cell death. Moreover, WA induced generation of reactive oxygen species (ROS) in PC-3 and DU-145, but not in normal fibroblasts. Immunocytochemistry and immuno-electron microscopy revealed that WA disrupted the vimentin cytoskeleton, possibly inducing the ROS generation, c-Fos expression and c-FLIP(L) suppression. These observations suggest that multiple events followed by disruption of the vimentin cytoskeleton play pivotal roles in WA-mediated cell death.