Abstract
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.
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Acknowledgements
We extend our sincere thanks to Ms. Peggy Donnelly and Ms. Denise Merrill for administrative support. AVK was supported by the Transdisciplinary Research on Energetics and Cancer (TREC) Training Program R25CA203650 (PI: Melinda Irwin).
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Conception and design of study were contributed by NV, AK. Data collection and analysis were contributed by NV, GP, AK. Preparation of figures was contributed by NV. Drafting and revising manuscript were contributed by NV, AK. NV, GP, AK approved final version of manuscript.
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Visavadiya, N.P., Pena, G.S. & Khamoui, A.V. Mitochondrial dynamics and quality control are altered in a hepatic cell culture model of cancer cachexia. Mol Cell Biochem 476, 23–34 (2021). https://doi.org/10.1007/s11010-020-03882-9
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DOI: https://doi.org/10.1007/s11010-020-03882-9