Myeloperoxidase expression in K-562 cells

Erich J. Baker, David Gerard, Carmen B. Lozzio, Albert T. Ichiki

Research output: Contribution to journalArticle

Abstract

Undifferentiated pluripotential K-562 cells maintain the capacity to differentiate along multiple hematopoietic lineages. Inducers of K-562 differentiation include sodium butyrate orhemin for hemoglobin synthesis, interferon-γ(IFN-γ) for the de novo expression of the MHC class I antigen, and phorbol 12-myristate 13-acetate (PMA) for the de novo synthesis of megakaryoblastic antigens CD41 and CD61. In spite of observed differentiation in response to these and other cytotoxic chemotherapeutic agents, K-562 cells do not traditionally demonstrate myeloperoxidase (MPO) biosynthesis or MPO mRNA constitutively or under various culture conditions. In this study, K-562 cells were cultured in HL-60 growth-conditioned medium (GCM) for up to 96 hours. MPO mRNA was transiently detected by RT-PCR techniques at 12, 24, and 48 hours. The de novo expression of MPO protein was subsequently detectable by intracellular flow cytometry at 24,48,72 and 96 hours. Immunogold staining and cytochemical analysis consequently demonstrated granularly-sequestered MPO in approximately 40% of HL-60 GCMcultured cells after 48 hours of culture. The sequential detection of myeloperoxidase mRNA and MPO biosynthesis is considered an indicator of serial maturation evocative of myeloblastic cells, and suggest that K-562 cells maintain the ability to differentiate along this lineage.

Original languageEnglish (US)
JournalBlood
Volume96
Issue number11 PART II
StatePublished - 2000

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Peroxidase
Biosynthesis
Messenger RNA
Platelet Membrane Glycoprotein IIb
Integrin beta3
Histocompatibility Antigens Class I
Butyric Acid
Flow cytometry
HL-60 Cells
Cytotoxins
Conditioned Culture Medium
Interferons
Cultured Cells
Flow Cytometry
Hemoglobins
Acetates
Staining and Labeling
Polymerase Chain Reaction
Growth
Proteins

All Science Journal Classification (ASJC) codes

  • Hematology

Cite this

Baker, E. J., Gerard, D., Lozzio, C. B., & Ichiki, A. T. (2000). Myeloperoxidase expression in K-562 cells. Blood, 96(11 PART II).

Myeloperoxidase expression in K-562 cells. / Baker, Erich J.; Gerard, David; Lozzio, Carmen B.; Ichiki, Albert T.

In: Blood, Vol. 96, No. 11 PART II, 2000.

Research output: Contribution to journalArticle

Baker, EJ, Gerard, D, Lozzio, CB & Ichiki, AT 2000, 'Myeloperoxidase expression in K-562 cells', Blood, vol. 96, no. 11 PART II.
Baker EJ, Gerard D, Lozzio CB, Ichiki AT. Myeloperoxidase expression in K-562 cells. Blood. 2000;96(11 PART II).
Baker, Erich J. ; Gerard, David ; Lozzio, Carmen B. ; Ichiki, Albert T. / Myeloperoxidase expression in K-562 cells. In: Blood. 2000 ; Vol. 96, No. 11 PART II.
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AB - Undifferentiated pluripotential K-562 cells maintain the capacity to differentiate along multiple hematopoietic lineages. Inducers of K-562 differentiation include sodium butyrate orhemin for hemoglobin synthesis, interferon-γ(IFN-γ) for the de novo expression of the MHC class I antigen, and phorbol 12-myristate 13-acetate (PMA) for the de novo synthesis of megakaryoblastic antigens CD41 and CD61. In spite of observed differentiation in response to these and other cytotoxic chemotherapeutic agents, K-562 cells do not traditionally demonstrate myeloperoxidase (MPO) biosynthesis or MPO mRNA constitutively or under various culture conditions. In this study, K-562 cells were cultured in HL-60 growth-conditioned medium (GCM) for up to 96 hours. MPO mRNA was transiently detected by RT-PCR techniques at 12, 24, and 48 hours. The de novo expression of MPO protein was subsequently detectable by intracellular flow cytometry at 24,48,72 and 96 hours. Immunogold staining and cytochemical analysis consequently demonstrated granularly-sequestered MPO in approximately 40% of HL-60 GCMcultured cells after 48 hours of culture. The sequential detection of myeloperoxidase mRNA and MPO biosynthesis is considered an indicator of serial maturation evocative of myeloblastic cells, and suggest that K-562 cells maintain the ability to differentiate along this lineage.

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