The regulation effect of liposomal transfection of antisense oligonucleotide on the alpha-globin in patients with severe beta-thalassemia / 中华血液学杂志
Chinese Journal of Hematology
; (12): 385-389, 2009.
Article
em Zh
| WPRIM
| ID: wpr-314475
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To study the effect of liposomal transfection of antisense oligonucleotide (ASON) on the erythroid cell alpha-globin gene in the patients with severe beta-thalassemia, and provide a new idea for beta-thalassemia gene therapy.</p><p><b>METHODS</b>A highly effective ASON targeting alpha-globin gene was transfected into severe beta-thalassemic erythroid cells cultured in vitro by liposomal at an optimal concentration. The expression level of alpha, beta, gamma-globin gene, the level of hemoglobin, and the excess alpha-globin chains precipitates in ASON group and control group were carefully analyzed by quantitative real-time PCR(Q-RT-PCR), high performance liquid chromatography (HPLC), and electron microscope, respectively.</p><p><b>RESULTS</b>The mRNA expression of alpha-globin gene was significantly lower in ASON group (9.04 +/- 0.29) than in control group (24.23 +/- 0.29) (P<0.01). Simultaneously, the disequilibrium between alpha- and beta-, gamma-globin gene expression was partly modified by ASON, the ratios of ASON group and control group being 0.79 +/- 0.02 and 2.26 +/- 0.06 respectively (P<0.01). HPLC demonstrated that the levels of HbA2 and HbF increased with downregulation of alpha-globin gene in beta-thalassemic erythroid cells, particularly HbF. The precipitates of alpha-globin chains in ASON group were lessened under electron microscope, particularly in early erythroblast while no change in the control group.</p><p><b>CONCLUSION</b>The high effective ASON contributes to inhibit the alpha-globin gene expression of severe beta-thalassemic erythroid cells, partly modify the disequilibrium between alpha-, beta- and gamma-globin gene expression and obviously reduce the precipitates of alpha-globin chains in erythroid cells. It might provide a new idea for gene therapy of beta-thalassemia.</p>
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Base de dados:
WPRIM
Assunto principal:
Terapêutica
/
Transfecção
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Terapia Genética
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Células Cultivadas
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Oligonucleotídeos Antissenso
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Talassemia beta
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Alfa-Globinas
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Globinas beta
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Gama-Globinas
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Genética
Limite:
Child
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Humans
Idioma:
Zh
Revista:
Chinese Journal of Hematology
Ano de publicação:
2009
Tipo de documento:
Article