摘要
目的探讨通过阻断BCL11A基因治疗重型β地中海贫血的可能性。方法对6例确诊重型β地中海贫血患者,CD34+免疫磁珠分离外周血CD34+胞后培养与诱导分化成体外原代红系细胞。培养第11天时处理组电转方法将针对BCL11A的siRNA电转入细胞,对照组导入随机siRNA,继续培养3天后以实时荧光定量检测XL和L型BCL11A水平,Western blot检测BCL11A表达水平。结果与对照组比较,BCL11A siRNA干扰组BCL11A mRNA表达水平下降0.48倍,γ-珠蛋白基因mRNA表达水平上升1.51倍。结论 BCL11A可作为重新激活HbF治疗β-地贫的分子靶标。
Objective To investigate the feasibility of treatment of thalssemia major by blocking BCL11A.Methods The CD34 ＋ cells were obtained from peripheral bood of 6 patients with β-thalassemia major with anti CD34 -tagged magnetic beads, and the combination of SCF, interleukin 3 （IL-3）, Epo and cyclosporine A were added to culture CD34＋ ceils for erythropoiesis differentiation and maturation into erythroblast. Small interfering RNA that target BCLI 1A was introduced into erythroblast with Gene Pulser at day 11, for control group random SiRNA was introduced.Western blot and quantity reverse transcriptase polymerase chain reaction （qRT-PCR） were performed to measure the expression level of BCLI 1A and γ-globin.Results Compared to control group, expression level of BCLllA mRNA decreased about 48%. Blocked expression of BCL11A was confirmed by Western blot.Gama-globin mRNA increased to 1.51 fold compared to control group.Conclusion BCLI 1A emerges as a therapeutic target for reactivation of HbF inβ-thalassemia.
出处
《华南国防医学杂志》
CAS
2014年第8期735-738,共4页
Military Medical Journal of South China
基金
广西壮族自治区自然科学基金项目(2011GXNSFA018196
2012GXNSFDA05301)
