[The regulation effect of liposomal transfection of antisense oligonucleotide on the alpha-globin in patients with severe beta-thalassemia].
OBJECTIVE
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Liu
Rong-Rong
RR
Department of Hematology, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.
Ma
Jie
J
Chen
Ping
P
Mo
Wu-Ning
WN
Lin
Wei-Xiong
WX
Lai
Yong-Rong
YR
D002478:Cells, Cultured; D002648:Child; D015316:Genetic Therapy; D006801:Humans; D008081:Liposomes; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D014162:Transfection; D055542:alpha-Globins / Q000235:genetics* / Q000378:metabolism; D055544:beta-Globins / Q000378:metabolism; D017086:beta-Thalassemia / Q000235:genetics / Q000378:metabolism* / Q000628:therapy; D055545:gamma-Globins / Q000378:metabolism
D008081:Liposomes; D016376:Oligonucleotides, Antisense; D055542:alpha-Globins; D055544:beta-Globins; D055545:gamma-Globins