title: Proteomic investigation of embryonic rat heart-derived H9c2 cell line sheds new light on the molecular phenotype of the popular cell model.
abstract: Due to their cardiac origin, H9c2 cells rank among the most popular cell lines in current cardiovascular research, yet molecular phenotype remains elusive. Hence, in this study we used proteomic approach to describe molecular phenotype of H9c2 cells in their undifferentiated (i.e., most frequently used) state, and its functional response to cardiotoxic drug doxorubicin. Of 1671 proteins identified by iTRAQ IEF/LC-MSMS analysis, only 12 proteins were characteristic for striated muscle cells and none was cardiac phenotype-specific. Targeted LC-SRM and western blot analyses confirmed that undifferentiated H9c2 cells are phenotypically considerably different to both primary neonatal cardiomyocytes and adult myocardium. These cells lack proteins essential for formation of striated muscle myofibrils or they express only minor amounts thereof. They also fail to express many proteins important for metabolism of muscle cells. The challenge with clinically relevant concentrations of doxorubicin did not induce a proteomic signature that has been previously noted in primary cardiomyocytes or adult hearts. Instead, several alterations previously described in other cells of mesodermal origin, such as fibroblasts, were observed (e.g., severe down-regulation of collagen synthesis pathway). In conclusion, the molecular phenotype of H9c2 cells resembles very immature myogenic cells with skeletal muscle commitment upon differentiation and thus, translatability of findings obtained in these cells deserves caution.
authors.affiliation: Department of Molecular Pathology and Biology, Faculty of Military Health Sciences, University of Defense, 500 01 Hradec Králové, Czech Republic.
authors.forename: Juraj
authors.initials: J
authors.lastname: Lenčo
authors.affiliation: Department of Pharmacology, Faculty of Medicine in Hradec Králové, Charles University in Prague, 500 38 Hradec Králové, Czech Republic.
authors.forename: Olga
authors.initials: O
authors.lastname: Lenčová-Popelová
authors.affiliation: Department of Molecular Pathology and Biology, Faculty of Military Health Sciences, University of Defense, 500 01 Hradec Králové, Czech Republic.
authors.forename: Marek
authors.initials: M
authors.lastname: Link
authors.affiliation: Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, 500 05 Hradec Králové, Czech Republic.
authors.forename: Anna
authors.initials: A
authors.lastname: Jirkovská
authors.affiliation: Biomedical Research Center, University Hospital Hradec Králové, 500 05 Hradec Králové, Czech Republic.
authors.forename: Vojtěch
authors.initials: V
authors.lastname: Tambor
authors.affiliation: Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, 500 05 Hradec Králové, Czech Republic.
authors.forename: Eliška
authors.initials: E
authors.lastname: Potůčková
authors.affiliation: Department of Molecular Pathology and Biology, Faculty of Military Health Sciences, University of Defense, 500 01 Hradec Králové, Czech Republic.
authors.forename: Jiří
authors.initials: J
authors.lastname: Stulík
authors.affiliation: Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, 500 05 Hradec Králové, Czech Republic.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Šimůnek
authors.affiliation: Department of Pharmacology, Faculty of Medicine in Hradec Králové, Charles University in Prague, 500 38 Hradec Králové, Czech Republic. Electronic address: sterbam@lfhk.cuni.cz.
authors.forename: Martin
authors.initials: M
authors.lastname: Štěrba
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000633:toxicity*; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D020543:Proteome / Q000032:analysis*; D051381:Rats; D013329:Structure-Activity Relationship
chemical_list: D020543:Proteome; D004317:Doxorubicin
keywords: Anthracyclines; Cardiomyocytes; Cell models; Doxorubicin; H9c2; Molecular phenotype; Proteomics