辅酶Q0（CoQ0）是从澳洲树芝（Antrodia camphorata，简称AC）中提取的衍生喹醌，具有抗癌活性。该研究检测了CoQ0（0-4 µM）对三阴性乳腺癌（MDA-MB-231和468）细胞中受抑制的抗EMT/转移和NLRP3炎症小体以及通过HIF-1α抑制改变Warburg效应的抗癌属性。使用MTT法、细胞迁移/侵袭实验、Western blotting、免疫荧光、代谢重编程和LC-ESI-MS评估了CoQ0的治疗潜力。CoQ0抑制了HIF-1α表达，并抑制了NLRP3炎症小体和ASC/半胱氨酸蛋白酶-1表达，接着降低了MDA-MB-231和468细胞中IL-1β和IL-18的表达。CoQ0通过降低CD44和增加CD24的表达来改善癌干细胞标记物。值得注意的是，CoQ0通过上调上皮标志物E-钙黏蛋白和下调间充质标志物N-钙黏蛋白来调节EMT。CoQ0抑制了葡萄糖摄取和乳酸积累。CoQ0还抑制了参与糖酵解的HIF-1α下游基因，如HK-2、LDH-A、PDK-1和PKM-2酶。CoQ0在正常氧和低氧（CoCl2）条件下，降低MDA-MB-231和468细胞中的胞外酸化率（ECAR）、糖酵解、糖酵解能力和糖酵解储备。CoQ0抑制了乳酸、FBP、2/3-PG和PEP等糖酵解中间代谢物和代谢产物的水平。CoQ0增加了正常氧和低氧（CoCl2）条件下的氧气消耗率（OCR）、基础呼吸、ATP产生、最大呼吸和备用容量。CoQ0增加了TCA循环代谢物，如柠檬酸、异柠檬酸和琥珀酸。CoQ0抑制了TNBC细胞的有氧糖酵解并增强了线粒体氧化磷酸化。在低氧条件下，CoQ0还减轻了MDA-MB-231和/或468细胞中的HIF-1α、GLUT1、糖酵解相关（HK-2、LDH-A和PFK-1）和转移相关（E-钙黏蛋白、N-钙黏蛋白和MMP-9）蛋白或mRNA表达。在LPS/ ATP刺激下，CoQ0抑制了NLRP3炎症小体/前半胱氨酸蛋白酶-1/IL-18激活和NFκB/iNOS表达。CoQ0还阻止了LPS/ ATP刺激的肿瘤迁移，并降低了LPS/ ATP刺激的N-钙黏蛋白和MMP-2/-9表达。本研究揭示，CoQ0抑制HIF-1α表达可能有助于抑制NLRP3介导的炎症、EMT/转移和三阴性乳腺癌的Warburg效应。©2023作者，独家许可Springer-Verlag GmbH Germany，Springer Nature的一部分。

Coenzyme Q0 (CoQ0) is a derivative quinone from Antrodia camphorata (AC) that exerts anticancer activities. This study examined the anticancer attributes of CoQ0 (0-4 µM) on inhibited anti-EMT/metastasis and NLRP3 inflammasome, and altered Warburg effects via HIF-1α inhibition in triple-negative breast cancer (MDA-MB-231 and 468) cells. MTT assay, cell migration/invasion assays, Western blotting, immunofluorescence, metabolic reprogramming, and LC-ESI-MS were carried out to assess the therapy potential of CoQ0. CoQ0 inhibited HIF-1α expression and suppressed the NLRP3 inflammasome and ASC/caspase-1 expression, followed by downregulation of IL-1β and IL-18 expression in MDA-MB-231 and 468 cells. CoQ0 ameliorated cancer stem-like markers by decreasing CD44 and increasing CD24 expression. Notably, CoQ0 modulated EMT by upregulating the epithelial marker E-cadherin and downregulating the mesenchymal marker N-cadherin. CoQ0 inhibited glucose uptake and lactate accumulation. CoQ0 also inhibited HIF-1α downstream genes involved in glycolysis, such as HK-2, LDH-A, PDK-1, and PKM-2 enzymes. CoQ0 decreased extracellular acidification rate (ECAR), glycolysis, glycolytic capacity, and glycolytic reserve in MDA-MB-231 and 468 cells under normoxic and hypoxic (CoCl2) conditions. CoQ0 inhibited the glycolytic intermediates lactate, FBP, and 2/3-PG, and PEP levels. CoQ0 increased oxygen consumption rate (OCR), basal respiration, ATP production, maximal respiration, and spare capacity under normoxic and hypoxic (CoCl2) conditions. CoQ0 increased TCA cycle metabolites, such as citrate, isocitrate, and succinate. CoQ0 inhibited aerobic glycolysis and enhanced mitochondrial oxidative phosphorylation in TNBC cells. Under hypoxic conditions, CoQ0 also mitigated HIF-1α, GLUT1, glycolytic-related (HK-2, LDH-A, and PFK-1), and metastasis-related (E-cadherin, N-cadherin, and MMP-9) protein or mRNA expression in MDA-MB-231 and/or 468 cells. Under LPS/ATP stimulation, CoQ0 inhibited NLRP3 inflammasome/procaspase-1/IL-18 activation and NFκB/iNOS expression. CoQ0 also hindered LPS/ATP-stimulated tumor migration and downregulated LPS/ATP-stimulated N-cadherin and MMP-2/-9 expression. The present study revealed that suppression of HIF-1α expression caused by CoQ0 may contribute to inhibition of NLRP3-mediated inflammation, EMT/metastasis, and Warburg effects of triple-negative breast cancers.© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.