Faculty of Medicine, University of Iceland, Reykjavik, Iceland.
Faculty of Pharmaceutical Sciences, University of Iceland, Reykjavik, Iceland.
Planta Med. 2022 Sep;88(11):891-898. doi: 10.1055/a-1579-6454. Epub 2021 Sep 14.
The lichen compound protolichesterinic acid (PA) has an anti-proliferative effect against several cancer cell lines of different origin. This effect cannot be explained by the known inhibitory activity of PA against 5- and 12-lipoxygenases. The aim was therefore to search for mechanisms for the anti-proliferative activity of PA. Two cancer cell lines of different origin, both sensitive to anti-proliferative effects of PA, were selected for this study, T-47D from breast cancer and AsPC-1 from pancreatic cancer. Morphological changes were assessed by transmission electron microscopy, HPLC coupled with TOF spectrometry was used for metabolomics, mitochondrial function was measured using the Agilent Seahorse XFp Real-time ATP assay and glucose/lactate levels by radiometry. Levels of glutathione, NADP/NADPH and reactive oxygen species [ROS] were measured by luminescence. Following exposure to PA both cell lines showed structural changes in mitochondria that were in line with a measured reduction in oxidative phosphorylation and increased glycolysis. These changes were more marked in T-47D, which had poorer mitochondrial function at baseline. PA was processed and expelled from the cells via the mercapturic pathway, which consumes glutathione. Nevertheless, glutathione levels were increased after 24 hours of exposure to PA, implying enhanced synthesis. Redox balance was not much affected and ROS levels were not increased. We conclude that PA is metabolically processed and expelled from cells, leading indirectly to increased glutathione levels with minimal effects on redox balance. The most marked effect was on mitochondrial structure and metabolic function implying that effects of PA may depend on mitochondrial fitness.
lichen 化合物原贝壳杉烯酸 (PA) 对多种不同来源的癌细胞系具有抗增殖作用。这种作用不能用 PA 对 5-和 12-脂氧合酶的已知抑制活性来解释。因此，目的是寻找 PA 抗增殖活性的机制。选择两种不同来源的对 PA 的抗增殖作用敏感的癌细胞系进行这项研究，一种是来自乳腺癌的 T-47D，另一种是来自胰腺癌的 AsPC-1。通过透射电子显微镜评估形态变化，高效液相色谱法与飞行时间质谱法耦合用于代谢组学，使用安捷伦 Seahorse XFp 实时 ATP 测定法测量线粒体功能，通过放射性测量法测量葡萄糖/乳酸水平。通过发光法测量谷胱甘肽、NADP/NADPH 和活性氧物种 [ROS] 的水平。PA 处理后，两种细胞系的线粒体均显示出结构变化，与氧化磷酸化减少和糖酵解增加一致。这些变化在 T-47D 中更为明显，T-47D 细胞的线粒体功能在基线时较差。PA 通过谷胱甘肽途径被细胞加工和排出，该途径消耗谷胱甘肽。然而，PA 暴露 24 小时后谷胱甘肽水平增加，表明合成增强。氧化还原平衡没有受到太大影响，ROS 水平也没有增加。我们得出结论，PA 在代谢上被加工并从细胞中排出，导致谷胱甘肽水平间接增加，氧化还原平衡几乎没有受到影响。最明显的影响是线粒体结构和代谢功能，这意味着 PA 的作用可能取决于线粒体的适应性。
