College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Gaoke Rd, Hangzhou, Zhejiang 311402, China.
College of Pharmaceutical Sciences, Zhejiang Chinese Medical University, Gaoke Rd, Hangzhou, Zhejiang 311402, China.
Phytomedicine. 2019 Apr;57:245-254. doi: 10.1016/j.phymed.2018.12.041. Epub 2018 Dec 31.
F3 is a novel fraction, for the first time isolated from Valeriana jatamansi Jones, which is a traditional Chinese folk medicine. Its anti-cancer potential and the underlying molecular mechanisms have not been well elucidated.
This study aims to investigate the anti-cancer effects of F3 on human breast cancer cell lines and its underlying mechanisms.
MTT assay was first performed to detect the effect of F3 on cell viability in human breast cancer cell lines and human mammary epithelial MCF-10A cells. Cell apoptosis, mitochondrial membrane potential and ROS level were detected by flow cytometry. Comet and immunofluorescence assays were utilized to assess DNA damage and expression of γ-H2AX. Autophagy were observed by AO staining and fluorescence microscopy. The expression of relative proteins was detected by western blotting. The xenograft model in nude mice was used to elucidate the effect of F3 on tumor growth and DNA damage in vivo.
F3 could significantly inhibit the growth of breast cancer cells in concentration-dependent manner by inducing apoptosis and has no obvious inhibitory effect of the growth on MCF-10A cells. Mechanistic studies demonstrated that F3-induced apoptosis was mediated by DNA damage as presented by DNA strand breaks and γ-H2AX activation that might be attacked by ROS accumulation. This triggered several key molecular events involving activation of MAPKs pathway. Further study showed that F3 induced autophagy with the autophagosome formation and increased LC3-II levels. Finally, in vivo study, F3 exhibited a potential antitumor effect and induced DNA damage in MDA-MB-231 xenografts.
The antitumorigenic activity of F3 was found in vitro and in vivo. These data suggest that F3 may be a potential natural active fraction for the treatment of human breast cancer.
F3 是一种新型的分数，首次从缬草（Valeriana jatamansi Jones）中分离出来，缬草是一种传统的中国民间药物。其抗癌潜力及其潜在的分子机制尚未得到充分阐明。
本研究旨在探讨 F3 对人乳腺癌细胞系的抗癌作用及其潜在机制。
首先通过 MTT 法检测 F3 对人乳腺癌细胞系和人乳腺上皮 MCF-10A 细胞活力的影响。通过流式细胞术检测细胞凋亡、线粒体膜电位和 ROS 水平。彗星和免疫荧光实验用于评估 DNA 损伤和 γ-H2AX 的表达。AO 染色和荧光显微镜观察自噬。通过 Western blot 检测相对蛋白的表达。裸鼠异种移植模型用于阐明 F3 对体内肿瘤生长和 DNA 损伤的影响。
F3 能够以浓度依赖的方式显著抑制乳腺癌细胞的生长，通过诱导细胞凋亡，对 MCF-10A 细胞的生长无明显抑制作用。机制研究表明，F3 诱导的细胞凋亡是由 DNA 损伤介导的，如 DNA 链断裂和 γ-H2AX 激活，这可能是由 ROS 积累引起的。这引发了几个关键的分子事件，涉及 MAPKs 途径的激活。进一步的研究表明，F3 诱导自噬，形成自噬体并增加 LC3-II 水平。最后，在体内研究中，F3 表现出潜在的抗肿瘤作用，并在 MDA-MB-231 异种移植瘤中诱导 DNA 损伤。
F3 在体外和体内均表现出抗肿瘤活性。这些数据表明，F3 可能是治疗人类乳腺癌的一种有潜力的天然活性成分。
