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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Drug evaluation based on phosphomimetic PDHA1 reveals the complexity of activity-related cell death in A549 non-small cell lung cancer cells

  • Jin, Ling (Department of Korean Medical Science, School of Korean Medicine, Pusan National University) ;
  • Cho, Minkyoung (Korean Medical Research Center for Healthy Aging, Pusan National University) ;
  • Kim, Bo-Sung (Department of Korean Medical Science, School of Korean Medicine, Pusan National University) ;
  • Han, Jung Ho (Department of Korean Medical Science, School of Korean Medicine, Pusan National University) ;
  • Park, Sungmi (Department of Internal Medicine, School of Medicine, Kyungpook National University) ;
  • Lee, In-Kyu (Department of Internal Medicine, School of Medicine, Kyungpook National University) ;
  • Ryu, Dongryeol (Department of Molecular Cell Biology, School of Medicine, Sungkyunkwan University) ;
  • Kim, Jae Ho (Department of Physiology, College of Medicine, Pusan National University) ;
  • Bae, Sung-Jin (Korean Medical Research Center for Healthy Aging, Pusan National University) ;
  • Ha, Ki-Tae (Department of Korean Medical Science, School of Korean Medicine, Pusan National University)
  • Received : 2021.07.26
  • Accepted : 2021.08.30
  • Published : 2021.11.30
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Abstract

Cancer cells predominantly generate energy via glycolysis, even in the presence of oxygen, to support abnormal cell proliferation. Suppression of PDHA1 by PDK1 prevents the conversion of cytoplasmic pyruvate into Acetyl-CoA. Several PDK inhibitors have been identified, but their clinical applications have not been successful for unclear reasons. In this study, endogenous PDHA1 in A549 cells was silenced by the CRISPR/Cas9 system, and PDHA1WT and PDHA13SD were transduced. Since PDHA13SD cannot be phosphorylated by PDKs, it was used to evaluate the specific activity of PDK inhibitors. This study highlights that PDHA1WT and PDHA13SD A549 cells can be used as a cell-based PDK inhibitor-distinction system to examine the relationship between PDH activity and cell death by established PDK inhibitors. Leelamine, huzhangoside A and otobaphenol induced PDH activity-dependent apoptosis, whereas AZD7545, VER-246608 and DCA effectively enhanced PDHA1 activity but little toxic to cancer cells. Furthermore, the activity of phosphomimetic PDHA1 revealed the complexity of its regulation, which requires further in-depth investigation.

Keywords

Acknowledgement

This work was supported by the National Research Foundation of Korea (NRF) grants funded by the Korean government (MIST; 2020R1C1C1003703 to Sung-Jin Bae, 2019R1A2C2003624 and 2021R1A4A1025662 to Ki-Tae Ha).

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