Lin Tzu-Ping, Chen Pei-Chun, Lin Ching-Yu, Wang Bi-Juan, Kuo Ying-Yu, Yeh Chien-Chih, Tseng Jen-Chih, Huo Chieh, Kao Cheng-Li, Shih Li-Jane, Chen Jen-Kun, Li Chia-Yang, Hour Tzyh-Chyuan, Chuu Chih-Pin
Faculty of Medicine, National Yang Ming Chiao Tung University, Hsinchu City, 30010, Taiwan.
Department of Urology, Taipei Veterans General Hospital, Taipei City, 11217, Taiwan.
BMC Cancer. 2025 Jan 16;25(1):95. doi: 10.1186/s12885-025-13477-6.
Caffeic acid phenethyl ester (CAPE) is the main bioactive component of poplar type propolis. We previously reported that treatment with caffeic acid phenethyl ester (CAPE) suppressed the cell proliferation, tumor growth, as well as migration and invasion of prostate cancer (PCa) cells via inhibition of signaling pathways of AKT, c-Myc, Wnt and EGFR. We also demonstrated that combined treatment of CAPE and docetaxel altered the genes involved in glycolysis and tricarboxylic acid (TCA) cycle. We therefore suspect that CAPE treatment may interfere glucose metabolism in PCa cells.
Seahorse Bioenergetics platform was applied to analyzed the extra cellular acidification rate (ECAR) and oxygen consumption rate (OCR) of PCa cells under CAPE treatment. UPLC-MSMS with Multiple Reaction Monitoring (MRM), PCR, and western blot were used to analyze the effects of CAPE on metabolites, genes, and proteins involved in glycolysis, TCA cycle and pentose phosphate pathway in PCa cells. Flow cytometry and ELISA were used to determine the level of reactive oxygen species in PCa cells being treated with CAPE.
Seahorse Bioenergetics analysis revealed that ECAR, glycolysis, OCR, and ATP production were elevated in C4-2B cells under CAPE treatment. Protein levels of glucose-6-phosphate dehydrogenase (G6PD), phosphogluconate dehydrogenase (PGD), glutaminase (GLS), phospho-AMPK Thr172 as well as abundance of pyruvate, lactate, ribulose-5-phosphate, and sedoheptulose-7-phosphate were increased in CAPE-treated C4-2B cells. ROS level decreased 48 h after treatment with CAPE. Co-treatment of AMPK inhibitor with CAPE exhibited additive growth inhibition on PCa cells.
Our study indicated that PCa cells attempted to overcome the CAPE-induced stress by upregulation of glycolysis and G6PD but failed to impede the growth inhibition caused by CAPE. Concurrent treatment of CAPE and inhibitors targeting glycolysis may be effective therapy for advanced PCa.
咖啡酸苯乙酯（CAPE）是杨树型蜂胶的主要生物活性成分。我们之前报道过，咖啡酸苯乙酯（CAPE）处理可通过抑制AKT、c-Myc、Wnt和EGFR信号通路来抑制前列腺癌细胞（PCa）的细胞增殖、肿瘤生长以及迁移和侵袭。我们还证明，CAPE与多西他赛联合处理会改变参与糖酵解和三羧酸（TCA）循环的基因。因此，我们怀疑CAPE处理可能会干扰PCa细胞中的葡萄糖代谢。
应用海马生物能量学平台分析CAPE处理下PCa细胞的细胞外酸化率（ECAR）和耗氧率（OCR）。采用多反应监测（MRM）的超高效液相色谱-质谱联用（UPLC-MSMS）、聚合酶链反应（PCR）和蛋白质印迹法分析CAPE对PCa细胞中参与糖酵解、TCA循环和磷酸戊糖途径的代谢物、基因和蛋白质的影响。采用流式细胞术和酶联免疫吸附测定（ELISA）法测定CAPE处理的PCa细胞中的活性氧水平。
海马生物能量学分析显示，CAPE处理下C4-2B细胞的ECAR、糖酵解、OCR和ATP生成均升高。CAPE处理的C4-2B细胞中，葡萄糖-6-磷酸脱氢酶（G6PD）、6-磷酸葡萄糖酸脱氢酶（PGD）、谷氨酰胺酶（GLS）、磷酸化腺苷酸活化蛋白激酶（AMPK）苏氨酸172的蛋白水平以及丙酮酸、乳酸、5-磷酸核酮糖和景天庚酮糖-7-磷酸的丰度均增加。CAPE处理48小时后，活性氧水平降低。AMPK抑制剂与CAPE联合处理对PCa细胞表现出相加的生长抑制作用。
我们的研究表明，PCa细胞试图通过上调糖酵解和G6PD来克服CAPE诱导的应激，但未能阻止CAPE引起的生长抑制。CAPE与靶向糖酵解的抑制剂联合处理可能是晚期PCa的有效治疗方法。
