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

Abstract

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 pharmacokinetic property and pharmacological activity of acteoside: A review

“…Additionally, AC up regulates the expression of Bax and caspase-3 and down regulates the expression of Bcl-2, leading to cell apoptosis. Furthermore, at the dose of 130 μM, AC increases TLR4 expression [71] . RNA metabolism regulation by microRNA or long noncoding RNA has been implicated in regulation of gene expression in cancer development [72] .…”

“… [69] 4T1 Inhibits the proliferative activity (IC 50 = 117 μM). [71] GBM cells Inhibits metastasis and promotes cell apoptosis and autophagy by let-7 g-5p/HMGA2/Wnt/β-catenin signaling. [73] Caco-2, HCT-116 Induces G1 cell cycle arrest and increases cell apoptosis by PI3K/AKT signaling.…”

The Anti-Tumor Efficacy of Verbascoside on Ovarian Cancer via Facilitating CCN1-AKT/NF-κB Pathway-Mediated M1 Macrophage Polarization

“…For subsequent experiments, SKOV3 and A2780 cells were treated with different VB concentrations (5,10,20,40, and 80 µM) and/or 20 µM LY294002. The concentrations of VB used in this study were decided according to previous studies (17,33,34). LY294002 is an AKT inhibitor that was used to block AKT/NF-kB pathway in this study (35).…”

Phytochemicals, Biological Activities, Molecular Mechanisms, and Future Prospects of Plantago asiatica L.

“…The apigenin ( 1 ) and luteolin ( 2 ) down-regulate NF-κB signaling (IKK, IκB, and NF-κB) and PI3K/AKT signaling (PI3K, AKT, and mTOR); activate or inactivate various signaling targets, such as Bcl-2, Bcl-xL, Bax, caspase-3, and caspase-9; and promote mitochondrial-mediated apoptosis, thereby producing anticancer effects. − Strangely, apigenin ( 1 ) can stimulate autophagic cell apoptosis of gastric cancer via inhibition of HIF-1α and Ezh2 under hypoxia conditions . Acteoside ( 27 ) increases the expression of Bax, caspase-3, and caspase-9 and reduces the expression of Bcl-2, cleaved caspase-3, and cleaved caspase-9 to promote tumor cell apoptosis. − In addition, plantamajoside ( 25 ) initiates cell apoptosis in lung cancer via dephosphorylation of MAPK and AKT, as well as mitochondria dysfunction …”