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Benzimidazoles induce concurrent apoptosis and pyroptosis of human glioblastoma cells via arresting cell cycle


Glioblastoma multiforme (GBM) is the most malignant and lethal primary brain tumor in adults accounting for about 50% of all gliomas. The only treatment available for GBM is the drug temozolomide, which unfortunately has frequent drug resistance issue. By analyzing the hub genes of GBM via weighted gene co-expression network analysis (WGCNA) of the cancer genome atlas (TCGA) dataset, and using the connectivity map (CMAP) platform for drug repurposing, we found that multiple azole compounds had potential anti-GBM activity. When their anti-GBM activity was examined, however, only three benzimidazole compounds, i.e. flubendazole, mebendazole and fenbendazole, potently and dose-dependently inhibited proliferation of U87 and U251 cells with IC50 values below 0.26 μM. Benzimidazoles (0.125−0.5 μM) dose-dependently suppressed DNA synthesis, cell migration and invasion, and regulated the expression of key epithelial-mesenchymal transition (EMT) markers in U87 and U251 cells. Benzimidazoles treatment also dose-dependently induced the GBM cell cycle arrest at the G2/M phase via the P53/P21/cyclin B1 pathway. Furthermore, the drugs triggered pyroptosis of GBM cells through the NF-κB/NLRP3/GSDMD pathway, and might also concurrently induced mitochondria-dependent apoptosis. In a nude mouse U87 cell xenograft model, administration of flubendazole (12.5, 25, and 50 mg · kg−1 · d−1, i.p, for 3 weeks) dose-dependently suppressed the tumor growth without obvious adverse effects. Taken together, our results demonstrated that benzimidazoles might be promising candidates for the treatment of GBM.

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Fig. 1: Drug repurposing based on hub gene identification by WGCNA analysis on the CMAP platform.
Fig. 2: Benzimidazoles inhibited proliferation of GBM cells in a dose- and time-dependent manner.
Fig. 3: Benzimidazoles inhibited migration and invasion of GBM cells in a dose-dependent manner.
Fig. 4: Benzimidazoles arrested GBM cell cycle at G2/M phase in a dose-dependent manner.
Fig. 5: Benzimidazoles triggered pyroptosis in GBM cells.
Fig. 6: Benzimidazoles induced mitochondria-dependent apoptosis of GBM cells.
Fig. 7: The effects of benzimidazoles were blocked by Z-VAD-FMK.
Fig. 8: Flubendazole suppressed the growth of GBM xenograft tumors.
Fig. 9: Schematic model for the mechanism of action of benzimidazoles.


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This work was supported by Beijing Natural Science Foundation (7212157), CAMS Innovation Fund for Medical Sciences (2016-I2M-3-007), National Natural Science Foundation of China (81803584, 81703536), Technology Major Projects for “Major New Drugs Innovation and Development” (2018ZX09711001-005-025, 2018ZX09711001-012).

Author information




GHD, JHW designed research; LWR, WL, XJZ, JYL, YHY performed research; LWR, SL, SZ, WQF and BX analyzed data; LWR wrote the paper.

Corresponding authors

Correspondence to Jin-hua Wang or Guan-hua Du.

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The authors declare no competing interests.

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Ren, Lw., Li, W., Zheng, Xj. et al. Benzimidazoles induce concurrent apoptosis and pyroptosis of human glioblastoma cells via arresting cell cycle. Acta Pharmacol Sin (2021).

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  • glioblastoma
  • benzimidazoles
  • cell cycle arrest
  • apoptosis
  • pyroptosis
  • drug repurposing


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