1. Increasing creatine kinase activity protects against hypoxia / reoxygenation injury but not against anthracycline toxicity in vitro
- Author
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Zervou, S, Whittington, HJ, Ostrowski, PJ, Cao, F, Tyler, J, Lake, HA, Neubauer, S, and Lygate, CA
- Subjects
lcsh:Medicine ,Apoptosis ,Bioenergetics ,Transfection ,Research and Analysis Methods ,Biochemistry ,Mice ,Open Reading Frames ,Gene Expression and Vector Techniques ,Medicine and Health Sciences ,Animals ,Humans ,Anthracyclines ,Cloning, Molecular ,Molecular Biology Techniques ,lcsh:Science ,Creatine Kinase ,Molecular Biology ,Energy-Producing Organelles ,Staining ,Molecular Biology Assays and Analysis Techniques ,Base Sequence ,Cell Death ,Organic Compounds ,Organic Chemistry ,lcsh:R ,Chemical Compounds ,Biology and Life Sciences ,Cell Staining ,Heart ,Cell Biology ,Creatine ,Cell Hypoxia ,Mitochondria ,Isoenzymes ,Oxygen ,Chemistry ,HEK293 Cells ,Cytoprotection ,Doxorubicin ,Cell Processes ,Specimen Preparation and Treatment ,Physical Sciences ,Cardiovascular Anatomy ,Hyperexpression Techniques ,lcsh:Q ,Cellular Structures and Organelles ,Anatomy ,Research Article - Abstract
The creatine kinase (CK) phosphagen system is fundamental to cellular energy homeostasis. Cardiomyocytes express three CK isoforms, namely the mitochondrial sarcomeric CKMT2 and the cytoplasmic CKM and CKB. We hypothesized that augmenting CK in vitro would preserve cell viability and function and sought to determine efficacy of the various isoforms. The open reading frame of each isoform was cloned into pcDNA3.1, followed by transfection and stable selection in human embryonic kidney cells (HEK293). CKMT2- CKM- and CKB-HEK293 cells had increased protein and total CK activity compared to non-transfected cells. Overexpressing any of the three CK isoforms reduced cell death in response to 18h hypoxia at 1% O2 followed by 2h re-oxygenation as assayed using propidium iodide: by 33% in CKMT2, 47% in CKM and 58% in CKB compared to non-transfected cells (P
- Published
- 2017