Identification of fetal hemoglobin-inducing agents using the human leukemia KU812 cell line

Sima Zein, Wei Li, Valya Ramakrishnan, Tzu Fang Lou, Sharanya Sivanand, Ashley Mackie, Betty Sue Pace

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Fetal hemoglobin (HbF) ameliorates the clinical severity of sickle cell disease; therefore continued research to identify efficacious HbF-inducing agents is desirable. In this study, we investigated KU812 leukemia cells that express the fetal γ-globin and adult β-globin genes, as a system for screening and discovery of novel HbF inducers. KU812 cells were analyzed in the presence or absence of fetal bovine serum and then expression levels of the globin genes, cell surface markers and transcription factors were quantified by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). For comparison, primary erythroid cells were grown in a two-phase liquid culture system. After drug inductions for 48-72 h, globin mRNA and HbF levels were quantified by RT-qPCR and enzyme-linked immunosorbent assay, respectively. Erythroid markers and transcription factors expression levels in KU812 cells were comparable to days 7-14 erythroid cells. We also tested several drugs including butyrate, trichostatin A, scriptaid, suberoylanilide hydroxamic acid and hydroxyurea, which induced γ-globin in KU812 cells; however, some agents also induced β-globin. A novel agent STI-571 was studied in the system, which non-selectively induced the globin genes. Additional studies showed comparable globin gene response patterns in KU812 and primary erythroid cells after treatments with the various drug inducers. Mechanisms of drug-mediated γ-globin induction in KU812 cells require signaling through the p38 mitogen-activated protein kinase pathway similar to that previously demonstrated in primary erythroid cells. These data suggest that KU812 cells serve as a good screening system to identify potential HbF inducers for the treatment of β-hemoglobinopathies.

Original languageEnglish (US)
Pages (from-to)1385-1394
Number of pages10
JournalExperimental Biology and Medicine
Volume235
Issue number11
DOIs
StatePublished - Nov 1 2010
Externally publishedYes

Fingerprint

Fetal Hemoglobin
Globins
Leukemia
Cells
Cell Line
Erythroid Cells
Genes
Polymerase chain reaction
Transcription
Pharmaceutical Preparations
Reverse Transcription
trichostatin A
Screening
Transcription Factors
Cell signaling
Hemoglobinopathies
Polymerase Chain Reaction
Immunosorbents
Hydroxyurea
Butyrates

Keywords

  • Fetal hemoglobin
  • KU812 cells
  • Primary erythroid cells
  • β-globin
  • γ-globin

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Identification of fetal hemoglobin-inducing agents using the human leukemia KU812 cell line. / Zein, Sima; Li, Wei; Ramakrishnan, Valya; Lou, Tzu Fang; Sivanand, Sharanya; Mackie, Ashley; Pace, Betty Sue.

In: Experimental Biology and Medicine, Vol. 235, No. 11, 01.11.2010, p. 1385-1394.

Research output: Contribution to journalArticle

Zein, Sima ; Li, Wei ; Ramakrishnan, Valya ; Lou, Tzu Fang ; Sivanand, Sharanya ; Mackie, Ashley ; Pace, Betty Sue. / Identification of fetal hemoglobin-inducing agents using the human leukemia KU812 cell line. In: Experimental Biology and Medicine. 2010 ; Vol. 235, No. 11. pp. 1385-1394.
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