siRNA抑制BCL11A表达对重型地中海贫血患者红系细胞γ-珠蛋白表达的影响  

Eeffect of SiRNA Tageting BCL11A onγ-globin Expression in β-thalassemia Erythroid Cells Cultured in vitro

作　　者：聂伟业 林万华[3] 罗瑞贵[2] 张新华[2] 尹晓林[2] 

机构地区：[1]桂林医学院研究生院,广西桂林541001 [2]解放军303医院血液科 [3]广西师范大学生命科学学院干细胞与医药生物技术实验室

出　　处：《华南国防医学杂志》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)

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.

关 键 词：重型Β地中海贫血 BCL11A基因 Γ基因 RNA干扰 

分 类 号：R566.7[医药卫生—呼吸系统]