毛蕊花糖苷对乳腺癌 4T1 细胞系的细胞毒性和凋亡作用。
The cytotoxicity and apoptotic effects of verbascoside on breast cancer 4T1 cell line.
机构信息
Student Research Center, Sabzevar University of Medical Sciences, Sabzevar, Iran.
Cellular and Molecular Research Center, Department of Physiology and Pharmacology, Faculty of Medicine, Sabzevar University of Medical Sciences, Sabzevar, Iran.
出版信息
BMC Pharmacol Toxicol. 2021 Nov 29;22(1):72. doi: 10.1186/s40360-021-00540-8.
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.
背景
尽管乳腺癌治疗取得了重大进展，但仍需要具有更低副作用的新型药物。在这方面，我们研究了毛蕊花糖苷在 4T1 小鼠乳腺肿瘤细胞中的抗癌特性。
方法
首先，用不同浓度（5-200μM 之间）的毛蕊花糖苷进行 MTT 测定，并计算 IC50。然后，评估处理后的 4T1 细胞中 Bax、Bcl-2 和 caspase-3 的表达。此外，我们还研究了 TLR4、MyD88 和 NF-κB 的表达，以确定毛蕊花糖苷抗增殖作用的潜在机制。此外，通过流式细胞术进行双 PI 和 Annexin V 检测，以确认毛蕊花糖苷诱导细胞凋亡的作用。
结果
MTT 测定结果显示，毛蕊花糖苷抑制 4T1 癌细胞的增殖（IC50 为 117μM），而对正常 HEK293T 细胞是安全的。通过 qRT-PCR，我们观察到毛蕊花糖苷处理（100、117 和 130μM）增加了 caspase-3 和 Bax 的表达，同时降低了 Bcl-2 的表达。此外，毛蕊花糖苷（100、117 和 130μM）仅在 130μM 剂量时增加了 TLR4 的表达，而增加了 MyD88 的表达，同时降低了 NF-κB 的表达。流式细胞术分析也证实毛蕊花糖苷在 117μM 时诱导 4T1 细胞凋亡。
结论
综上所述，我们的数据表明毛蕊花糖苷对正常细胞是一种安全的天然化合物，而对 4T1 细胞具有通过 TLR4 轴诱导细胞凋亡的作用。
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