Hydroxyurea generates nitric oxide in human erythroid cells: Mechanisms for c-globin gene activation

Tzu Fang Lou, Manisha Singh, Ashley Mackie, Wei Li, Betty S. Pace

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Hydroxyurea (HU) induces fetal hemoglobin synthesis through activation of cyclic guanine monophosphate (cGMP) signaling. Studies in sickle cell patients demonstrated increased circulating nitric oxide (NO) levels after oral HU treatment. However, the direct measurement of NO in erythroid cells and its role in fetal hemoglobin induction have not been defined. Therefore, we quantified the level of nitrate and nitrite (NOx) generated by HU in human erythroid progenitors in the presence of three nitric oxide synthase inhibitors (NOS), including NG-monomethyl-L-arginine (L-NMMA). In addition, cGMP levels were measured in the presence or absence of the pathway inhibitor 1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one, which blocks soluble guanylyl cyclase formation. HU treatment increased NOx levels and γ-globin transcription in K562 and primary erythroid cells, which was augmented when HU was combined with L-NMMA. Pretreatment with the cGMP pathway inhibitor reversed γ-gene activation by HU. These data demonstrate the direct stimulation of cellular NO and cGMP signaling in erythroid progenitors by HU as a possible mechanism for γ-globin gene activation.

Original languageEnglish (US)
Pages (from-to)1374-1382
Number of pages9
JournalExperimental Biology and Medicine
Volume234
Issue number11
DOIs
StatePublished - Nov 1 2009

Fingerprint

Erythroid Cells
Hydroxyurea
Globins
Transcriptional Activation
Nitric Oxide
Genes
Chemical activation
Guanine
omega-N-Methylarginine
Fetal Hemoglobin
Guanylate Cyclase
Transcription
Nitrites
Nitric Oxide Synthase
Nitrates
Therapeutics

Keywords

  • Fetal hemoglobin
  • Hydroxyurea
  • Nitric oxide
  • Sickle-cell disease
  • cGMP

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Hydroxyurea generates nitric oxide in human erythroid cells : Mechanisms for c-globin gene activation. / Lou, Tzu Fang; Singh, Manisha; Mackie, Ashley; Li, Wei; Pace, Betty S.

In: Experimental Biology and Medicine, Vol. 234, No. 11, 01.11.2009, p. 1374-1382.

Research output: Contribution to journalArticle

Lou, Tzu Fang ; Singh, Manisha ; Mackie, Ashley ; Li, Wei ; Pace, Betty S. / Hydroxyurea generates nitric oxide in human erythroid cells : Mechanisms for c-globin gene activation. In: Experimental Biology and Medicine. 2009 ; Vol. 234, No. 11. pp. 1374-1382.
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