Background: Triple-negative breast cancer (TNBC) is one of the most prevalent neoplastic diseases worldwide, but efficacious treatments for this pathological condition are still challenging. The lack of an effective targeted therapy also leads to a poor prognosis for patients affected by TNBC. In the present study, we repurposed the distinctive inhibitory effects of flubendazole, a traditional anthelmintic drug, towards the putative modulation of proliferation and migration of TNBC in vitro and in vivo./nMethods: According to a series of experimental approaches, including immunofluorescence (IF), immunoblotting (IB), siRNA and GFP-mRFP-LC3 plasmid transfection, respectively, we have found that flubendazole is capable of inducing autophagic cell death and apoptosis, thus exerting some anti-proliferative and anti-migration activity in TNBC cells. The therapeutic effects of flubendazole were evaluated by xenograft mouse models, followed by immunohistochemistry (IHC), IF and IB. Changes in the gene expression profiles of flubendazole-treated TNBC cells were analyzed by RNA sequencing (RNA-seq) and validated by IB. The potential binding mode of flubendazole and EVA1A was predicted by molecular docking and demonstrated by site-directed mutagenesis./nResults: We have presently found that flubendazole exhibits a considerable anti-proliferative activity in vitro and in vivo. Mechanistically, the induction of autophagic cell death appears to be pivotal for flubendazole-mediated growth inhibition of TNBC cells, whereas blocking autophagy was able to improve the survival rate and migration ability of flubendazole-treated TNBC cells. Specifically, RNA-seq analysis showed that flubendazole treatment could promote the up-regulation of EVA1A. Flubendazole may regulate autophagy and apoptosis by targeting EVA1A, thus affecting the mechanisms of TNBC proliferation and migration. Furthermore, Thr113 may be the key amino acid residues for the binding of flubendazole to EVA1A./nConclusion: Our results provide novel insights towards the putative anti-cancer efficacy of flubendazole. Furthermore, here we show that flubendazole could serve as a potential therapeutic drug in TNBC. Altogether, this study highlights the possibility of this repurposed autophagic inducer for future cancer treatments.

中文翻译：

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氟苯达唑通过靶向EVA1A调节的自噬和三阴性乳腺癌中的细胞凋亡引发抗癌作用。

背景：三阴性乳腺癌（TNBC）是世界上最普遍的赘生性疾病之一，但是对于这种病理状况的有效治疗仍然具有挑战性。缺乏有效的靶向治疗还导致受TNBC影响的患者预后较差。在本研究中，我们改变用途氟苯咪唑，传统的驱虫药的独特的抑制作用，对增殖和TNBC迁移的推定的调制在体外和体内./n方法：根据一系列实验方法，分别包括免疫荧光（IF），免疫印迹（IB），siRNA和GFP-mRFP-LC3质粒转染，我们发现flubendazole能够诱导自噬细胞死亡和凋亡，从而发挥一些抗TNBC细胞中的增殖和抗迁移活性。通过异种移植小鼠模型，随后免疫组织化学（IHC），IF和IB评估了氟苯达唑的治疗效果。氟苯达唑处理的TNBC细胞基因表达谱的变化通过RNA测序（RNA-seq）分析并通过IB验证。的氟苯咪唑和EVA1A潜在结合模式是通过分子对接预测与通过位点定向mutagenesis./n证实结果：我们目前发现氟苯达唑在体外和体内均表现出相当大的抗增殖活性。从机制上讲，自噬细胞死亡的诱导似乎是对flubendazole介导的TNBC细胞生长抑制的关键，而阻断自噬则能够提高flubendazole处理的TNBC细胞的存活率和迁移能力。具体来说，RNA-seq分析表明，氟苯达唑处理可以促进EVA1A的上调。氟苯达唑可能通过靶向EVA1A来调节自噬和凋亡，从而影响TNBC增殖和迁移的机制。此外，Thr113可以是氨基为对氟苯咪唑EVA1A./n的结合残基的键结论：我们的结果为氟苯达唑的假定抗癌功效提供了新颖的见解。此外，在这里我们显示了氟苯达唑可以作为TNBC中潜在的治疗药物。总而言之，这项研究突出了这种重新定位的自噬诱导剂用于未来癌症治疗的可能性。