Abstract：
OBJECTIVE: Dual-Interventions targeting glucose and oxidative metabolism are receiving increasing attention in cancer therapy. Sorafenib (S) and Metformin (M), two gold-standards in liver cancer, are known for their mitochondrial inhibitory capacity. Fasting, a glucose-limiting strategy, is also emerging as chemotherapy adjuvant. Herein, we explore the anti-carcinogenic response of nutrient restriction in combination with sorafenib:metformin (NR-S:M).
RESULTS: Our data demonstrates that, independently of liver cancer aggressiveness, fasting synergistically boosts the anti-proliferative effects of S:M co-treatment. Metabolic and Cellular plasticity was determined by the examination of mitochondrial and glycolytic activity, cell cycle modulation, activation of cellular apoptosis, and regulation of key signaling and metabolic enzymes. Under NR-S:M conditions, early apoptotic events and the pro-apoptotic Bcl-xS/Bcl-xL ratio were found increased. NR-S:M induced the highest retention in cellular SubG1 phase, consistent with the presence of DNA fragments from cellular apoptosis. Mitochondrial functionality, Mitochondrial ATP-linked respiration, Maximal respiration and Spare respiratory capacity, were all found blunted under NR-S:M conditions. Basal Glycolysis, Glycolytic reserve, and glycolytic capacity, together with the expression of glycogenic (PKM), gluconeogenic (PCK1 and G6PC3), and glycogenolytic enzymes (PYGL, PGM1, and G6PC3), were also negatively impacted by NR-S:M. Lastly, a TMT-proteomic approach corroborated the synchronization of liver cancer metabolic reprogramming with the activation of molecular pathways to drive a quiescent-like status of energetic-collapse and cellular death.
CONCLUSIONS: Altogether, we show that the energy-based polytherapy NR-S:M blunts cellular, metabolic and molecular plasticity of liver cancer. Notwithstanding the in vitro design of this study, it holds a promising therapeutic tool worthy of exploration for this tumor pathology.
RESULTS: Our data demonstrates that, independently of liver cancer aggressiveness, fasting synergistically boosts the anti-proliferative effects of S:M co-treatment. Metabolic and Cellular plasticity was determined by the examination of mitochondrial and glycolytic activity, cell cycle modulation, activation of cellular apoptosis, and regulation of key signaling and metabolic enzymes. Under NR-S:M conditions, early apoptotic events and the pro-apoptotic Bcl-xS/Bcl-xL ratio were found increased. NR-S:M induced the highest retention in cellular SubG1 phase, consistent with the presence of DNA fragments from cellular apoptosis. Mitochondrial functionality, Mitochondrial ATP-linked respiration, Maximal respiration and Spare respiratory capacity, were all found blunted under NR-S:M conditions. Basal Glycolysis, Glycolytic reserve, and glycolytic capacity, together with the expression of glycogenic (PKM), gluconeogenic (PCK1 and G6PC3), and glycogenolytic enzymes (PYGL, PGM1, and G6PC3), were also negatively impacted by NR-S:M. Lastly, a TMT-proteomic approach corroborated the synchronization of liver cancer metabolic reprogramming with the activation of molecular pathways to drive a quiescent-like status of energetic-collapse and cellular death.
CONCLUSIONS: Altogether, we show that the energy-based polytherapy NR-S:M blunts cellular, metabolic and molecular plasticity of liver cancer. Notwithstanding the in vitro design of this study, it holds a promising therapeutic tool worthy of exploration for this tumor pathology.
摘要：
目的：针对葡萄糖和氧化代谢的双重干预在癌症治疗中越来越受到重视。索拉非尼（S）和二甲双胍（M），肝癌的两个黄金标准，以其线粒体抑制能力而闻名。禁食,葡萄糖限制策略，也正在成为化疗的辅助药物。在这里，我们探讨了营养限制联合索拉非尼：二甲双胍（NR-S：M）的抗癌反应。
结果：我们的数据表明，独立于肝癌的侵袭性，禁食协同增强S：M联合治疗的抗增殖作用。通过检查线粒体和糖酵解活性来确定代谢和细胞可塑性，细胞周期调制，细胞凋亡的激活，以及关键信号和代谢酶的调节。在NR-S:M条件下，发现早期凋亡事件和促凋亡Bcl-xS/Bcl-xL比值增加.NR-S：M诱导细胞亚G1期的最高保留，与来自细胞凋亡的DNA片段的存在一致。线粒体功能，线粒体ATP相关呼吸，最大呼吸和备用呼吸能力，在NR-S：M条件下都被发现钝化。基础糖酵解,糖酵解储备，和糖酵解能力，连同糖原（PKM）的表达，糖异生（PCK1和G6PC3），和糖原分解酶(PYGL，PGM1和G6PC3),也受到NR-S:M的负面影响最后,TMT-蛋白质组学方法证实了肝癌代谢重编程与分子途径激活的同步，以驱动能量崩溃和细胞死亡的静止状态。
结论：总而言之，我们表明，基于能量的综合疗法NR-S：M钝化细胞，肝癌的代谢和分子可塑性。尽管本研究的体外设计，它为这种肿瘤病理提供了一个值得探索的有前途的治疗工具。
结果：我们的数据表明，独立于肝癌的侵袭性，禁食协同增强S：M联合治疗的抗增殖作用。通过检查线粒体和糖酵解活性来确定代谢和细胞可塑性，细胞周期调制，细胞凋亡的激活，以及关键信号和代谢酶的调节。在NR-S:M条件下，发现早期凋亡事件和促凋亡Bcl-xS/Bcl-xL比值增加.NR-S：M诱导细胞亚G1期的最高保留，与来自细胞凋亡的DNA片段的存在一致。线粒体功能，线粒体ATP相关呼吸，最大呼吸和备用呼吸能力，在NR-S：M条件下都被发现钝化。基础糖酵解,糖酵解储备，和糖酵解能力，连同糖原（PKM）的表达，糖异生（PCK1和G6PC3），和糖原分解酶(PYGL，PGM1和G6PC3),也受到NR-S:M的负面影响最后,TMT-蛋白质组学方法证实了肝癌代谢重编程与分子途径激活的同步，以驱动能量崩溃和细胞死亡的静止状态。
结论：总而言之，我们表明，基于能量的综合疗法NR-S：M钝化细胞，肝癌的代谢和分子可塑性。尽管本研究的体外设计，它为这种肿瘤病理提供了一个值得探索的有前途的治疗工具。
