School of Engineering and Material Science, Queen Mary University of London, Mile End Road, London E1 4NS, UK.
Int J Mol Sci. 2020 Jun 23;21(12):4450. doi: 10.3390/ijms21124450.
The mechanical state of cells is a critical part of their healthy functioning and it is controlled primarily by cytoskeletal networks (actin, microtubules and intermediate filaments). Drug-based strategies targeting the assembly of a given cytoskeletal network are often used to pinpoint their role in cellular function. Unlike actin and microtubules, there has been limited interest in the role of intermediate filaments, and fewer drugs have thus been identified and characterised as modulators of its assembly. Here, we evaluate whether Withaferin-A (WFA), an established disruptor of vimentin filaments, can also be used to modulate keratin filament assembly. Our results show that in keratinocytes, which are keratin-rich but vimentin-absent, Withaferin-A disrupts keratin filaments. Importantly, the dosages required are similar to those previously reported to disrupt vimentin in other cell types. Furthermore, Withaferin-A-induced keratin disassembly is accompanied by changes in cell stiffness and migration. Therefore, we propose that WFA can be repurposed as a useful drug to disrupt the keratin cytoskeleton in epithelial cells.
细胞的机械状态是其正常功能的关键部分，主要由细胞骨架网络（肌动蛋白、微管和中间丝）控制。基于药物的策略，针对特定细胞骨架网络的组装，常用于确定其在细胞功能中的作用。与肌动蛋白和微管不同，人们对中间丝的作用的兴趣有限，因此，被鉴定为其组装调节剂的药物也较少。在这里，我们评估了 Withaferin-A（一种已被证实能破坏中间丝的药物）是否也可用于调节角蛋白丝的组装。我们的结果表明，在富含角蛋白但不含波形蛋白的角质细胞中，Withaferin-A 破坏了角蛋白丝。重要的是，所需的剂量与以前在其他细胞类型中报道的破坏波形蛋白的剂量相似。此外，Withaferin-A 诱导的角蛋白解聚伴随着细胞硬度和迁移的变化。因此，我们提出 WFA 可以被重新用作一种有用的药物，以破坏上皮细胞中的角蛋白细胞骨架。
