Abstract
Breast cancer is one of the most malignant tumors and is associated with high mortality rates among women. Lycium barbarum polysaccharide (LBP) is an extract from the fruits of the traditional Chinese herb, L. barbarum. LBP is a promising anticancer drug, due to its high activity and low toxicity. Although it has anticancer properties, its mechanisms of action have not been fully established. Ferroptosis, which is a novel anticancer strategy, is a cell death mechanism that relies on iron-dependent lipid reactive oxygen species (ROS) accumulation. In this study, human breast cancer cells (Michigan Cancer Foundation-7 (MCF-7) and MD Anderson-Metastatic Breast-231 (MDA-MB-231)) were treated with LBP. LBP inhibited their viability and proliferation in association with high levels of ferroptosis. Therefore, we aimed to ascertain whether LBP reduced cell viability through ferroptosis. We found that the structure and function of mitochondria, lipid peroxidation, and expression of solute carrier family 7 member 11 (SLC7A11, also known as xCT, the light-chain subunit of cystine/glutamate antiporter system Xc−) and glutathione peroxidase 4 (GPX4) were altered by LBP. Moreover, the ferroptosis inhibitor, Ferrostatin-1 (Fer-1), rescued LBP-induced ferroptosis-associated events including reduced cell viability and glutathione (GSH) production, accumulation of intracellular free divalent iron ions and malondialdehyde (MDA), and down-regulation of the expression of xCT and GPX4. Erastin (xCT inhibitor) and RSL3 (GPX4 inhibitor) inhibited the expression of xCT and GPX4, respectively, which was lower after the co-treatment of LBP with Erastin and RSL3. These results suggest that LBP effectively prevents breast cancer cell proliferation and promotes ferroptosis via the xCT/GPX4 pathway. Therefore, LBP exhibits novel anticancer properties by triggering ferroptosis, and may be a potential therapeutic option for breast cancer.
摘要
目的
乳腺癌是女性高发的恶性肿瘤之一。枸杞多糖(Lycium barbarumpolysaccharide, LBP)作为我国传统名贵中药材枸杞子的主要活性成分, 因其高活性和低毒性逐渐成为抗癌药物的新选择。已有研究表明LBP对多种肿瘤细胞的生长具有抑制作用, 但机制尚不明确。铁死亡是一种新的细胞死亡方式, 主要依赖铁的脂质过氧化, 引发细胞死亡, 而LBP与铁死亡的关系尚未见明确报道。因此, 本研究旨在探索LBP是否通过诱导铁死亡发挥抗癌作用。
创新点
我们首次发现LBP通过蛋白溶质载体家族7成员11和谷胱甘肽过氧化酶4(xCT/GPX4)途径介导乳腺癌细胞铁死亡, 这对深入理解LBP治疗乳腺癌的中药药理机制具有重要意义, 也为乳腺癌的中药抗癌研究提供了新思路。
方法
通过细胞活性检测, 确定LBP可抑制人乳腺癌细胞MCF-7和MDA-MB-231的存活与增殖。为了探索其作用机制, 本研究利用转录组测序(RNA-seq)检测LBP处理前后对MCF-7细胞的影响, 结果发现LBP处理后, 差异基因富集到铁死亡信号通路。后续通过对LBP处理后铁死亡标志物(线粒体形态、线粒体膜电位和脂质过氧化水平等)的检测进一步验证转录组结果。通过加入铁死亡抑制剂Fer-1, 研究LBP对两种乳腺癌的细胞存活率、氧化应激水平和胞内自由二价铁的影响, 探究LBP是否通过诱导铁死亡介导抗癌作用, 最后, 通过检测两种乳腺癌细胞中铁死亡调控蛋白xCT/GPX4的表达水平明确具体其作用机制。
结论
LBP通过xCT/GPX4信号通路诱导铁死亡, 并降低乳腺癌细胞的存活与增殖。研究结果揭示LBP通过铁死亡途径展现出新的抗癌特性, 可能成为乳腺癌的潜在治疗选择。
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (No. 81960480) and the Key Research and Development Program of Ningxia, China (No. 2018BEB04008).
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Xing DU, Jingjing ZHANG, Ling LIU, Bo XU, Hang HAN, Wenjie DAI, and Xiuying PEI performed the experimental research and data analysis. Xing DU, Shaozhang HOU, and Xufeng FU contributed to the study design, data analysis, and writing and editing of the manuscript. All authors have read and approved the final manuscript, and therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Xing DU, Jingjing ZHANG, Ling LIU, Bo XU, Hang HAN, Wenjie DAI, Xiuying PEI, Xufeng FU, and Shaozhang HOU declare that they have no conflict of interest.
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Du, X., Zhang, J., Liu, L. et al. A novel anticancer property of Lycium barbarum polysaccharide in triggering ferroptosis of breast cancer cells. J. Zhejiang Univ. Sci. B 23, 286–299 (2022). https://doi.org/10.1631/jzus.B2100748
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DOI: https://doi.org/10.1631/jzus.B2100748