Granger D L, Taintor R R, Cook J L, Hibbs J B
J Clin Invest. 1980 Feb;65(2):357-70. doi: 10.1172/JCI109679.
Cytotoxic activated macrophages (CM) inhibited the growth of neoplastic L1210 cells in vitro but L1210 cell death was minimal to nonexistent. L1210 cells injured by CM were separated from macrophages and studied in an isolated system. CM-injured L1210 cells had an absolute requirement for glucose or another glycolyzable hexose (mannose or fructose) for at least 40 h after removal from macrophages. If the culture medium lacked sufficient concentration of one of these sugars, CM-injured L1210 cells died within 4 h. Uninjured L1210 cells cultured alone or with peptone-stimulated macrophages had no such requirement and maintained complete viability in hexoseless medium. The hexose requirement of CM-injured L1210 cells could not be fulfilled by other naturally occurring monosaccharides, glucose or mannose derivatives, or substrates that can be oxidized by mitochondria. The concentration requirements for glucose, mannose, and fructose by CM-injured L1210 cells correlated with the concentrations required to support maximal glycolysis of these sugars by other murine ascites cells. A concentration of 2-deoxy-D-glucose which completely inhibited L1210 cell glycolysis also complete prevented the ability of glucose or mannose to maintain viability of CM-injured L1210 cells. Interaction with CM led to inhibition of L1210 cell mitochondrial oxidative phosphorylation. This was supported by the findings that: (a) CM-injured L1210 cells had no Pasteur effect; their rate of aerobic glycolysis was the same as the rate of anaerobic glycolysis of uninjured L1210 cells, (b) Endogenous respiration of CM-injured L1210 cells was 15% of normal. Maximal inhibition of uninjured L1210 cell respiration by a specific mitochondrial poison (oligomycin) was nearly the same (13% of normal). It followed that CM-injured L1210 cells required hexose for chemical energy production via the glycolytic pathway. CM-induced mitochondrial injury occurred in five other neoplastic cell lines tested.
细胞毒性活化巨噬细胞（CM）在体外抑制肿瘤性L1210细胞的生长，但L1210细胞的死亡微乎其微甚至不存在。将受CM损伤的L1210细胞与巨噬细胞分离，并在一个孤立系统中进行研究。从巨噬细胞中分离出来后，受CM损伤的L1210细胞在至少40小时内绝对需要葡萄糖或另一种可糖酵解的己糖（甘露糖或果糖）。如果培养基中缺乏这些糖之一的足够浓度，受CM损伤的L1210细胞会在4小时内死亡。单独培养或与蛋白胨刺激的巨噬细胞一起培养的未受损L1210细胞没有这种需求，并且在无己糖培养基中保持完全活力。受CM损伤的L1210细胞对己糖的需求不能通过其他天然存在的单糖、葡萄糖或甘露糖衍生物，或可被线粒体氧化的底物来满足。受CM损伤的L1210细胞对葡萄糖、甘露糖和果糖的浓度需求与其他小鼠腹水细胞支持这些糖最大糖酵解所需的浓度相关。完全抑制L1210细胞糖酵解的2-脱氧-D-葡萄糖浓度也完全阻止了葡萄糖或甘露糖维持受CM损伤的L1210细胞活力的能力。与CM的相互作用导致L1210细胞线粒体氧化磷酸化受到抑制。以下发现支持了这一点：（a）受CM损伤的L1210细胞没有巴斯德效应；它们的有氧糖酵解速率与未受损L1210细胞的无氧糖酵解速率相同，（b）受CM损伤的L1210细胞的内源性呼吸是正常的15%。特定线粒体毒物（寡霉素）对未受损L1210细胞呼吸的最大抑制作用几乎相同（正常的13%）。由此可见，受CM损伤的L1210细胞需要己糖通过糖酵解途径产生化学能量。CM诱导的线粒体损伤在测试的其他五种肿瘤细胞系中也有发生。
