Flubendazole elicits anti-cancer effects via targeting EVA1A-modulated autophagy and apoptosis in Triple-negative Breast Cancer

Abstract

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 . Methods: 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. Results: 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. Conclusion: 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.

PNO1 regulates autophagy and apoptosis of hepatocellular carcinoma via the MAPK signaling pathway

“…Several studies have shown that autophagy may occur upstream of apoptosis 29 , 30 . To determine whether PNO1-mediated autophagy is related to apoptosis in HCC cells, we used rapamycin and 3-MA to activate and inhibit autophagy, respectively.…”

“…The relationship between autophagy and apoptosis is known to be complex. Accumulating evidence shows that autophagy and apoptosis usually occur within the same cell, and autophagy may serve as upstream regulation mechanism of apoptosis 29 , 30 , 54 . According to the different interactions of autophagy and apoptosis in tumor, it could be divided into synergistic, promoting, and antagonistic effects 37 , 55 .…”

EVA1B to Evaluate the Tumor Immune Microenvironment and Clinical Prognosis in Glioma

“…In humans, the EVA1 family includes three main groups: EVA1A, EVA1B, and EVA1C. Previous studies have revealed that EVA1A, an endoplasmic reticulum-related protein were involved in regulating autophagy and apoptosis-related mechanisms in many cancers (11,12). Additionally, Bang-Yi Lin et al found that EVA1A promotes papillary thyroid cancer progression and epithelial-mesenchymal transition via upregulating immunerelated signaling pathways (11).…”

Progress of Breast Cancer basic research in China

“…Flubendazole, a broad-spectrum anthelmintic drug belonging to benzimidazole group, has been repurposed as a promising anti-cancer agent 94 . Zhen et al revealed flubendazole induced autophagic cell death by targeting Eva-1 homolog A (EVA1A) and suppressed TNBC proliferation and migration 95 . Moreover, resistomycin, QN-1 and emodin attenuates TNBC progression via binding to Pellino-1 to promote FBXO11-mediated SNAIL/SLUG degradation 96 , down-regulating c-MYC transcription 97 and reducing VEGFA transcription 98 , respectively.…”