Sulforaphane-N-Acetyl-Cysteine inhibited autophagy leading to apoptosis via Hsp70-mediated microtubule disruption
Sulforaphane-N-Acetyl-Cysteine inhibited autophagy leading to apoptosis via Hsp70-mediated microtubule disruption
复制标题
萝卜硫素-N-乙酰半胱氨酸抑制自噬，通过 Hsp70 介导的微管破坏导致细胞凋亡
DOI：
10.1016/j.canlet.2018.05.035
复制
发表时间：
2018-01-01
期刊：
影响因子：
9.7
通讯作者：
Wu, Wei
中科院分区：
文献类型：
--
作者：
Hu, Yabin;Zhou, Yan;Wu, Wei
莱菔硫烷-N-乙酰半胱氨酸（SFN-NAC）是一种潜在的抑制人非小细胞肺癌（NSCLC）的药物，但其作用机制尚不清楚。在这里，我们发现SFN-NAC诱导细胞凋亡，通过流式细胞仪分析和透射电镜。此外，SFN-NAC增加LC 3 II/LC 3 I和LC 3斑点的数量，但Western blot显示SFN-NAC抑制细胞自噬响应于巴弗洛霉素Al和SFN-NAC的共同处理。此外，免疫荧光染色和Western印迹显示SFN-NAC通过下调α-微管蛋白和磷酸化ERK 1/2介导的Stathmin-1触发微管破坏，引起细胞凋亡。此外，SFN-NAC通过磷酸化ERK 1/2上调Hsp 70。共聚焦显微镜和免疫沉淀实验表明，SFN-NAC促进了Hsp 70和α-tubulin的共定位和相互作用; Hsp 70的敲除增强了SFN-NAC诱导的微管断裂，降低了LC 3 II/LC 3 I，促进了细胞凋亡。有趣的是，组织微阵列分析显示，α-微管蛋白或Hsp 70的表达增加与NSCLC恶性分级相关，表明微管和Hsp 70是SFN-NAC的两个关键靶点。这些结果将使我们对SFN-NAC诱导的细胞凋亡有一个新的认识，以便我们开发更有效的治疗NSCLC的疗法。
Sulforaphane-N-acetyl-cysteine (SFN-NAC) is a potential drug to inhibit human non-small cell lung cancer (NSCLC), but the underlying mechanisms are elusive. Here, we uncovered that SFN-NAC induced apoptosis via flow cytometer assay and transmission electron microscopy. Further, SFN-NAC increased LC3 II/LC3 I and the number of LC3 punctas, but Western blot showed that SFN-NAC inhibited cell autophagy in response to a co treatment of Bafilomycin Al and SFN-NAC. Furthermore, immunofluorescence staining and Western blot showed that SFN-NAC triggered microtubule disruption causing apoptosis via downregulating a-tubulin and phosphorylated ERK1/2-mediated Stathmin-1. Besides, SFN-NAC upregulated Hsp70 via phosphorylating ERK1/2. Confocal microscopy and immunoprecipitation assay showed that SFN-NAC promoted the colocalization and interaction of Hsp70 and alpha-tubulin; knockdown of Hsp70 enhanced SFN-NAC-induced microtubule disruption, lowered LC3 II/LC3 I and promoted apoptosis. Interestingly, tissue microarray analysis showed that the increased expression of either alpha-tubulin or Hsp70 correlated to NSCLC malignant grading, indicating that microtubule and Hsp70 are two key targets for SFN-NAC. These results will give us a new insight into SFN-NAC-induced apoptosis so that we develop more efficient therapeutics to treat NSCLC.