TY - JOUR
T1 - Characterization of K562 cells
T2 - uncovering novel chromosomes, assessing transferrin receptor expression, and probing pharmacological therapies
AU - Karagiannis, Tom C.
AU - Wall, Meaghan
AU - Ververis, Katherine
AU - Pitsillou, Eleni
AU - Tortorella, Stephanie M.
AU - Wood, Peter A.
AU - Rafehi, Haloom
AU - Khurana, Ishant
AU - Maxwell, Scott S.
AU - Hung, Andrew
AU - Vongsvivut, Jitraporn
AU - El-Osta, Assam
N1 - Funding Information:
The authors would like to acknowledge the use of the facilities provided by Monash Micro Imaging at AMREP and particularly, the expert assistance from Drs Stephen Cody and Iśka Carmichael. Also we acknowledge the technical expertise of Dr. Clement Khaw (Singapore Bioimaging Consortium-Nikon Imaging Centre) for N-SIM super resolution imaging and Dr Darren Henstrigde (Cellular and Molecular metabolism, Baker Heart and Diabetes Institute) for assistance with the Seahorse XF96 extracellular flux analyzer. M.W. was supported by the Leukaemia Foundation of Australia. EP is supported by an Australian Government Research Training Program Scholarship. We thank the National Computing Infrastructure (NCI), and the Pawsey Supercomputing Centre in Australia (funded by the Australian Government). Further, we thank the Spartan High Performance Computing service (University of Melbourne), and the Partnership for Advanced Computing in Europe (PRACE) for awarding the access to Piz Daint, hosted at the Swiss National Supercomputing Centre (CSCS), Switzerland. Supported in part by the Victorian Government’s Operational Infrastructure Support Program.
Funding Information:
The authors would like to acknowledge the use of the facilities provided by Monash Micro Imaging at AMREP and particularly, the expert assistance from Drs Stephen Cody and Iśka Carmichael. Also we acknowledge the technical expertise of Dr. Clement Khaw (Singapore Bioimaging Consortium-Nikon Imaging Centre) for N-SIM super resolution imaging and Dr Darren Henstrigde (Cellular and Molecular metabolism, Baker Heart and Diabetes Institute) for assistance with the Seahorse XF96 extracellular flux analyzer. M.W. was supported by the Leukaemia Foundation of Australia. EP is supported by an Australian Government Research Training Program Scholarship. We thank the National Computing Infrastructure (NCI), and the Pawsey Supercomputing Centre in Australia (funded by the Australian Government). Further, we thank the Spartan High Performance Computing service (University of Melbourne), and the Partnership for Advanced Computing in Europe (PRACE) for awarding the access to Piz Daint, hosted at the Swiss National Supercomputing Centre (CSCS), Switzerland. Supported in part by the Victorian Government’s Operational Infrastructure Support Program.
Publisher Copyright:
© 2023, Crown.
PY - 2023/9
Y1 - 2023/9
N2 - Human erythroleukemic K562 cells represent the prototypical cell culture model of chronic myeloid leukemia (CML). The cells are pseudo-triploid and positive for the Philadelphia chromosome. Therefore, K562 cells have been widely used for investigating the BCR/ABL1 oncogene and the tyrosine kinase inhibitor, imatinib-mesylate. Further, K562 cells overexpress transferrin receptors (TfR) and have been used as a model for targeting cytotoxic therapies, via receptor-mediated endocytosis. Here, we have characterized K562 cells focusing on the karyotype of cells in prolonged culture, regulation of expression of TfR in wildtype (WT) and doxorubicin-resistant cells, and responses to histone deacetylase inhibition (HDACi). Karyotype analysis indicates novel chromosomes and gene expression analysis suggests a shift of cultured K562 cells away from patient-derived leukemic cells. We confirm the high expression of TfR on K562 cells using immunofluorescence and cell-surface receptor binding radioassays. Importantly, high TfR expression is observed in patient-derived cells, and we highlight the persistent expression of TfR following doxorubicin acquired resistance. Epigenetic analysis indicates that permissive histone acetylation and methylation at the promoter region regulates the transcription of TfR in K562 cells. Finally, we show relatively high expression of HDAC enzymes in K562 cells and demonstrate the chemotoxic effects of HDACi, using the FDA-approved hydroxamic acid, vorinostat. Together with a description of morphology, infrared spectral analysis, and examination of metabolic properties, we provide a comprehensive characterization of K562 cells. Overall, K562 cell culture systems remain widely used for the investigation of novel therapeutics for CML, which is particularly important in cases of imatinib-mesylate resistance. Graphical abstract: [Figure not available: see fulltext.].
AB - Human erythroleukemic K562 cells represent the prototypical cell culture model of chronic myeloid leukemia (CML). The cells are pseudo-triploid and positive for the Philadelphia chromosome. Therefore, K562 cells have been widely used for investigating the BCR/ABL1 oncogene and the tyrosine kinase inhibitor, imatinib-mesylate. Further, K562 cells overexpress transferrin receptors (TfR) and have been used as a model for targeting cytotoxic therapies, via receptor-mediated endocytosis. Here, we have characterized K562 cells focusing on the karyotype of cells in prolonged culture, regulation of expression of TfR in wildtype (WT) and doxorubicin-resistant cells, and responses to histone deacetylase inhibition (HDACi). Karyotype analysis indicates novel chromosomes and gene expression analysis suggests a shift of cultured K562 cells away from patient-derived leukemic cells. We confirm the high expression of TfR on K562 cells using immunofluorescence and cell-surface receptor binding radioassays. Importantly, high TfR expression is observed in patient-derived cells, and we highlight the persistent expression of TfR following doxorubicin acquired resistance. Epigenetic analysis indicates that permissive histone acetylation and methylation at the promoter region regulates the transcription of TfR in K562 cells. Finally, we show relatively high expression of HDAC enzymes in K562 cells and demonstrate the chemotoxic effects of HDACi, using the FDA-approved hydroxamic acid, vorinostat. Together with a description of morphology, infrared spectral analysis, and examination of metabolic properties, we provide a comprehensive characterization of K562 cells. Overall, K562 cell culture systems remain widely used for the investigation of novel therapeutics for CML, which is particularly important in cases of imatinib-mesylate resistance. Graphical abstract: [Figure not available: see fulltext.].
KW - BCR-ABL
KW - Chronic myeloid leukemia
KW - Doxorubicin
KW - Histone deacetylase inhibitors
KW - K562 cells
KW - Philadelphia chromosome
KW - Transferrin receptors
UR - http://www.scopus.com/inward/record.url?scp=85168066677&partnerID=8YFLogxK
U2 - 10.1007/s00018-023-04905-6
DO - 10.1007/s00018-023-04905-6
M3 - Article
C2 - 37578596
AN - SCOPUS:85168066677
SN - 1420-682X
VL - 80
JO - Cellular and Molecular Life Sciences
JF - Cellular and Molecular Life Sciences
IS - 9
M1 - 248
ER -