The anthelmintic drug flubendazole induces cell apoptosis and inhibits NF-κB signaling in esophageal squamous cell carcinoma

Abstract

Background and objectivesThe nuclear factor kappa B (NF-κB) signaling is activated in esophageal squamous cell carcinoma (ESCC) and can be used as a potential target for anti-ESCC drug discovery. In this study, we aimed to investigate the function of flubendazole as a novel NF-κB inhibitor in ESCC cells.Materials and methodsCell Counting Kit-8 assay was carried out to assess cell viability of ESCC cells. Flow cytometry and immunoblotting were performed to examine cell apoptosis. Immunoblotting assay was used to analyze the protein expression of NF-κB signaling. Luciferase assay was performed to explore the activation of NF-κB. Plasmids were transfected into ESCC cells using Lipofectamine® 2000.ResultsIn this study, the anthelmintic drug flubendazole was found to inhibit the activation of IκBα kinases (IKKs), block the activation of IκBα, and decrease the phosphorylation of NF-κB p65, which could be a novel NF-κB inhibitor in ESCC cells. We also found that flubendazole inhibited the cell survival of different ESCC cells and induced cell apoptosis in both EC9706 and TE1 cells. Moreover, overexpression of constitutively activated IKKβ markedly decreased the cytotoxic effect of flubendazole on EC9706 and TE1 cells. In addition, flubendazole also showed a synergistic effect on ESCC cells when combined with doxorubicin.ConclusionThe results above demonstrated that flubendazole showed its anti-tumor action by suppressing the NF-κB signaling pathway and suggested that flubendazole might be re-purposed for anti-ESCC therapy in clinic as a single agent or in combination with other anti-tumor drugs.

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

“…In tumor cells, flubendazole has been recently shown to affect microtubule dynamics, inhibit angiogenesis and cell metastasis, as well as to induce mitotic catastrophe and apoptosis 15 - 24 . Mechanistically, flubendazole may also target NF-κB signaling, which is required for esophageal squamous carcinoma cell survival 17 . In colorectal and breast cancer cells, flubendazole exerts anti-tumor and anti-metastatic activities by inhibiting STAT3 22 , 26 .…”

Cytotoxic Effects of Flubendazole in Human Lung Cancer Cell Line A549

“…Flubendazole is another anthelminthic drug belonging to the benzimidazole family with anticancer effects by inhibiting microtubules function (11,22). In a study, Michalis et al (2015) examined the flubendazole effect on several cell lines of neuroblastoma cancer and introduced it as a potential anti-neuroblastoma compound.…”

“…We used six repetitions for each group, while replication was considered as a standard template. Flubendazole solution was prepared at concentrations of 1, 2, 5, and 10 µM in DMSO and diluted with a medium, then filtered and stored at 4˚C (11,12).…”

Ubiquitin‐specific protease 44 inhibits cell growth by suppressing AKT signaling in non‐small cell lung cancer

“…A549 and H1975 cells were stably infected with PLVX‐NC or PLVX‐USP44 lentivirus, and then cells were incubated in 96‐well plates for 0, 1, 3, or 5 days, followed by Cell Counting Kit‐8 (CCK‐8) staining according to the manufacturer's instructions (Biotool, Houston, TX) as described previously …”

“…A549 and H1975 cells were stably infected with PLVX-NC or PLVX-USP44 lentivirus, and then cells were incubated in 96-well plates for 0, 1, 3, or 5 days, followed by Cell Counting Kit-8 (CCK-8) staining according to the manufacturer's instructions (Biotool, Houston, TX) as described previously. 19,20 2.7 | Cell cycle analysis A549 and H1975 cells were infected with lentivirus-delivered PLVX-NC or PLVX-USP44 for 4 days before cell cycle analysis. Subsequently, cells were harvested and fixed with 70% cold ethanol overnight.…”