Hepatic mitochondrial function loss is associated with cancer cachexia pathology in vivo. Here, we examined if hepatic mitochondrial defects observed in vivo in the cachexic liver also recapitulate during the in vitro treatment of mouse hepatocytes with tumor conditioned media. In vitro experiments were combined with proteome-wide expression analysis of cachexic liver tissue curated for mitochondrial dynamics and quality control proteins, to determine the fidelity of hepatic mitochondrial maladaptation in cancer cachexia pathology. AML12 hepatocytes were exposed to colon-26 (C26) and Lewis lung carcinoma (LLC) conditioned media for 6–72 h and assayed for cell viability, membrane potential, respiratory function, H₂O₂ production, total ROS/RNS, and mitochondrial dynamics and quality control proteins by immunoblotting. Liver tissue from cachexic C26 mice was analyzed by TMT-based quantitative proteomics for in vivo comparison. Cell viability, membrane potential, H₂O₂ production, total ROS/RNS, and respiration were decreased 48–72 h after exposure to C26 and/or LLC. Protein expression of treated hepatocytes and cachexic liver tissue showed altered mitochondrial dynamics and quality control, in a manner that suggests limited fusion and content mixing, but also impaired ability to fragment and clear damaged mitochondria. Two strategies to maintain mitochondrial health, therefore, may not be functioning sufficiently in the cachexic liver. Together these findings imply adverse effects of C26 and LLC exposure on hepatocyte health, due to impaired mitochondrial function and remodeling. Exposure of mouse hepatocytes to tumor conditioned media models aspects of cachexic liver mitochondria dysfunction in vivo and validates the importance of hepatic mitochondrial maladaptation in cancer cachexia pathology.

肝线粒体功能丧失与体内癌症恶病质病理相关。在这里，我们检查了在体内用恶病质肝观察到的肝线粒体缺陷在用肿瘤条件培养基体外治疗小鼠肝细胞的过程中是否也重现了。体外实验与针对线粒体动力学和质量控制蛋白精心挑选的恶病质肝组织的全蛋白质组表达分析相结合，以确定在癌症恶病质病理中肝线粒体适应不良的保真度。将AML12肝细胞暴露于结肠26（C26）和Lewis肺癌（LLC）条件培养基中6–72 h，并检测其细胞活力，膜电位，呼吸功能，H ₂ O ₂生产，总ROS / RNS，线粒体动力学和质量控制蛋白（通过免疫印迹）。通过基于TMT的定量蛋白质组学分析来自恶病质C26小鼠的肝组织，以进行体内比较。细胞活力，膜电位，H ₂ O ₂暴露于C26和/或LLC后48-72小时，生产，总ROS / RNS和呼吸减少。处理过的肝细胞和恶病质肝组织的蛋白质表达表明线粒体动力学和质量控制发生了变化，这表明融合和含量混合受到限制，但破坏和清除受损线粒体的能力也受到损害。因此，两种维持线粒体健康的策略在恶病质肝脏中可能无法充分发挥作用。由于线粒体功能和重塑受损，这些发现共同暗示了C26和LLC暴露对肝细胞健康的不利影响。将小鼠肝细胞暴露于肿瘤条件培养基可模拟体内恶病质肝线粒体功能障碍的各个方面，并验证了肝线粒体适应不良在癌症恶病质病理学中的重要性。