Abstract 1382: Expression and intracellular translocation of cancer biomarkers in hepatocarcinoma cells induced by changes in mitochondrial metabolism

Abstract

Hepatocarcinoma is one of the most common cancer types in the U.S. with limited treatment options and poor survival rate. Recently, cancer bioenergetics emerged as a promising source of targets to develop novel anticancer agents. The identification of biomarkers is essential for screening and validation of small molecules proposed as chemotherapeutic agents. In this regard, there is a great need for high-throughput/high-content methods, which require short and reproducible sample preparation. In this project, we used the erastin-like compound X1 in the well-characterized SNU-449 hepatocarcinoma cell line. Erastin initially enhances mitochondrial function and ROS formation leading to subsequent mitochondrial dysfunction. As readout, we employed directly and indirectly labeled fluorescent antibodies to visualize and quantify expression of proliferative and antiproliferative markers Ki-67 and cytoskeleton-associated protein 4/p63 (CKAP4/p63), as well as expression and intracellular translocation of the tumor suppressor protein p53. Using the Agilent BioTek Cytation 5 cell imaging multimode reader and Agilent BioTek Gen5 microplate reader and imager software, we successfully validated these markers and the corresponding antibodies as tools to study long-term effects of xenobiotics on mitochondrial bioenergetics. The method allows for fast and reliable screening of potential drug candidates. Acknowledgement: This work was supported in part by the Chan Zuckerberg Initiative, Silicon Valley Community Foundation, and the NIH S10 OD028663 to M.G. and by the U.S. National Institutes of Health (NIH/NCI) R01 CA184456, and South Carolina Translational Research Pilot Project UL1 TR001450-SCTR to E.N.M. Citation Format: Bradley R. Larson, Monika Gooz, Eduardo Maldonado. Expression and intracellular translocation of cancer biomarkers in hepatocarcinoma cells induced by changes in mitochondrial metabolism [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1382.

Cancer Bioenergetics and Tumor Microenvironments—Enhancing Chemotherapeutics and Targeting Resistant Niches through Nanosystems

“…Studies in eight different cancer cell lines and patients' plasma samples have elucidated the characteristics of cfDNA, confirming its release exclusively by cancer cells, with different release patterns [211][212][213][214][215][216]. Screening of the bioenergetics flux parameters showed a correlation between cfDNA release patterns and cellular origin, cancer status, and proliferation phase [217][218][219][220]. A significant dependence correlating with cfDNA release was noted with aerobic glycolysis but not OXPHOS [219,221,222].…”

“…Hence, the significance of cfDNA in cancer diagnosis and precision medicine is underscored by its potential and routine use in the clinical diagnosis and management of several cancers. TME-associated molecular, cellular, stromal, and signaling mechanisms are restructured by mitochondrial energy modulations to facilitate growth and resistance to therapy [214,220]. Hence, TME-associated tumor vasculature, immune cells, and stromal cells recruited to the tumor in all development and proliferative phases can be manipulated to manage cancer by altering or halting tumor advancement [199].…”

“…Hence, TME-associated tumor vasculature, immune cells, and stromal cells recruited to the tumor in all development and proliferative phases can be manipulated to manage cancer by altering or halting tumor advancement [199]. Metabolic derangement within the TME leads to a complex increase in gasotransmitters (nitric oxide, carbon monoxide and hydrogen sulfide), and metabolites such as lactic acid, K + , and kynurenine [220,228,229]. These molecules alter the pH of TME in a resource-constraint environment, repress immune functions, and help tumor growth.…”