重型β地贫患儿珠蛋白肽链失衡的调控研究

Regulation research on globin peptide imbalance in children with severe β-thalassemia

目的研究重型β地贫患儿外周血珠蛋白α、β、γmRNA比例变化及miR-144基因对其平衡的调控作用。方法分离培养患者外周血早红细胞,真核表达质粒PmiR-144-luc转染后培养72h,运用RT-PCR分别检测重型β地贫患儿空白组、基因转染组与健康对照组外周血α、β、γmRNA的比率变化。结果与健康对照组比较,重型β地贫患儿外周血珠蛋白α/β、α/β+γ、γ/β+γmRNA的比例水平均升高,差别有统计学意义;与重型β地贫患儿空白组比较,基因转染组珠蛋白α/β、α/β+γmRNA的比例水平均降低,差别有统计学意义;而γ/β+γmRNA的比例水平变化无统计意义。结论 miR-144基因可能调控重型β地贫患儿珠蛋白α、β、γmRNA基因比例的变化,将为重型β地贫的基因调节治疗提供新的实验依据。

Objective To detect mRNA expression levels of globin peptide α,β and Tin the peripheral blood of children with se vere β-thalassemia, and analyze the relationship between miR 144 gene and the changes of globin ratio. Methods The early red blood cells were isolated from children with severe β-thalassemia and in vitro cultured, and changes of the ratio of α ,β and 7 mRNA were detected by quantitative RT PCR for β thalassemia group,miR-144 gene transfection group and the healthy control group, 72 hours after the transfection of eukaryotic expression plasmids PmiR-144-luc into the isolated early red blood cells. Results Com pared with the healthy control group, the ratios of globin α ,β,α /β＋γ and α/β＋γ mRNA in peripheral blood of children with severe thalassemia were elevated, with statistical significance. Compared with ~ thalassemia control group, the ratios of α /β and β＋γ mRNA in gene transfection group were reduced, with statistical difference, but the ratio of β＋γ mRNA was not with any statis tical difference. Conclusion miR-144 might take part in the regulation of the ratio of α ,β and γ globin gene expression in children with severe γ-thalassemia, and this finding might provide a new experimental basis for gene regulation treatment of severe β thalassemia.