|
|
 Previous Article | Table of Contents | Next Article 
Blood, Vol. 95 No. 3 (February 1), 2000:
pp. 863-869
Quantitative PCR analysis of HbF inducers in primary human
adult erythroid cells
Reginald D. Smith,
Jin Li,
Constance T. Noguchi, and
Alan N. Schechter
From the Laboratory of Chemical Biology, National Institute of
Diabetes and Digestive and Kidney Diseases, National Institutes of
Health, Bethesda, MD 20892.
The development and evaluation of drugs to elevate fetal hemoglobin
in the treatment of the genetic diseases of hemoglobin would be
facilitated by the availability of reliable cell assays. We have used
real-time, quantitative polymerase chain reaction (PCR) analyses of
globin messenger RNA (mRNA) levels in a biphasic, erythropoietin-dependent primary culture system for human adult erythroid cells in order to assay compounds for their ability to
modulate levels of adult ( ) and fetal ( ) globin mRNA.
Complementary DNA synthesized from total RNA extracted at timed
intervals from aliquots of cells were assayed throughout the period
that the culture was studied. -globin mRNA levels were found to be
much lower (less than 1%) than -globin mRNA levels. At
concentrations of agents chosen for minimal effect on cell division, we
find that the 3 drugs studied, 5-azacytidine
(5µmol/L), hydroxyurea (40µmol/L),
and butyric acid (0.5mmol/L), significantly increase -globin mRNA levels. Interestingly, hydroxyurea also had a small stimulatory effect on -globin mRNA levels, while butyric acid caused
a twofold inhibition of -globin mRNA levels, and 5-azacytidine had
little effect on -globin mRNA levels. The net result of all 3 drugs
was to increase the /( + ) mRNA ratios by threefold to
fivefold. These data suggest that the mechanism is distinct for each
drug. The profile of butyric-acid-induced changes on globin gene
expression is also quite distinct from changes produced by trichostatin
A, a known histone deacetylase inhibitor. Quantitative PCR analyses of
human erythroid cells should prove useful for studying the mechanism(s)
of action of known inducers of -globin and identifying new drug candidates.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
V. P. Cokic, S. A. Andric, S. S. Stojilkovic, C. T. Noguchi, and A. N. Schechter
Hydroxyurea nitrosylates and activates soluble guanylyl cyclase in human erythroid cells
Blood,
February 1, 2008;
111(3):
1117 - 1123.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Kleinbongard, R. Schulz, T. Rassaf, T. Lauer, A. Dejam, T. Jax, I. Kumara, P. Gharini, S. Kabanova, B. Ozuyaman, et al.
Red blood cells express a functional endothelial nitric oxide synthase
Blood,
April 1, 2006;
107(7):
2943 - 2951.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. C. Tang, J. Zhu, W. Liu, K. Chin, J. Sun, L. Chen, J. A. Hanover, and G. P. Rodgers
The hydroxyurea-induced small GTP-binding protein SAR modulates {gamma}-globin gene expression in human erythroid cells
Blood,
November 1, 2005;
106(9):
3256 - 3263.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. P. Frade, D. W. Warnock, and B. A. Arthington-Skaggs
Rapid Quantification of Drug Resistance Gene Expression in Candida albicans by Reverse Transcriptase LightCycler PCR and Fluorescent Probe Hybridization
J. Clin. Microbiol.,
May 1, 2004;
42(5):
2085 - 2093.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. Wataganara, E. S. LeShane, A. Y. Chen, L. Borgatta, I. Peter, K. L. Johnson, and D. W. Bianchi
Plasma {gamma}-Globin Gene Expression Suggests that Fetal Hematopoietic Cells Contribute to the Pool of Circulating Cell-Free Fetal Nucleic Acids during Pregnancy
Clin. Chem.,
April 1, 2004;
50(4):
689 - 693.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
O. Witt, S. Monkemeyer, G. Ronndahl, B. Erdlenbruch, D. Reinhardt, K. Kanbach, and A. Pekrun
Induction of fetal hemoglobin expression by the histone deacetylase inhibitor apicidin
Blood,
March 1, 2003;
101(5):
2001 - 2007.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
U. Wojda, P. Noel, and J. L. Miller
Fetal and adult hemoglobin production during adult erythropoiesis: coordinate expression correlates with cell proliferation
Blood,
April 15, 2002;
99(8):
3005 - 3013.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Samakoglu, E. Fattori, S. Lamartina, C. Toniatti, D. Stockholm, J. M. Heard, and D. Bohl
{beta}Minor-globin messenger RNA accumulation in reticulocytes governs improved erythropoiesis in {beta} thalassemic mice after erythropoietin complementary DNA electrotransfer in muscles
Blood,
April 15, 2001;
97(8):
2213 - 2220.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. D. Smith, J. D. Malley, and A. N. Schechter
Quantitative analysis of globin gene induction in single human erythroleukemic cells
Nucleic Acids Res.,
December 15, 2000;
28(24):
4998 - 5004.
[Abstract]
[Full Text]
[PDF]
|
|
|
| |
