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Background  Despite significant advancements in breast cancer therapy, novel drugs with lower side effects are still being demanded. In this regard, we investigated the anti-cancer features of verbascoside in 4 T1 mouse mammary tumor cell.    Methods  First, MTT assay was performed with various concentrations (ranging between 5 to 200 μM) of verbascoside and IC50 was calculated. Then the expression of  Bax ,  Bcl-2 , and  caspase-3  was evaluated in treated 4 T1 cells. In addition, we investigated the expression of  TLR4 ,  MyD88 , and  NF-κB  to ascertain the underlying mechanism of the anti-proliferative feature of verbascoside. Also, flow cytometry followed by double PI and Annexin V was conducted to confirm the apoptosis-inducing effect of verbascoside.    Results  Our results from MTT assay showed verbascoside inhibits proliferation of 4 T1 cancer cells (IC50 117 μM) while is safe for normal HEK293T cells. By qRT-PCR, we observed that verbascoside treatment (100, 117 and, 130 μM) increases the expression of  caspase-3  and  Bax  while reduces the expression of  Bcl-2 . Also, verbascoside (100, 117 and, 130 μM) increased the expression of  TLR4  only at 130 μM dose and the expression of  MyD88  whereas reduced the expression of  NF-κB  at mRNA level. Flow cytometry analysis also confirmed verbascoside induces apoptosis in 4 T1 cells at 117 μM.    Conclusion  Taken together, our data showed verbascoside is a safe natural compound for normal cells while has apoptosis-inducing feature through TLR4 axis on 4 T1 cells.

The cytotoxicity and apoptotic effects of verbascoside on breast cancer 4T1 cell line