RNA干扰对体外培养HUVECs增殖和CD105表达的影响  

Effect of Silencing CD105 Gene with siRNA on Proliferation of HUVECs and Expression of CD105

作　　者：丁怡[1,2] 张明昌[2] 

机构地区：[1]中国人民解放军广州军区武汉总医院眼科,武汉430070 [2]华中科技大学同济医学院附属协和医院眼科,武汉430022

出　　处：《华中科技大学学报（医学版）》2011年第6期662-665,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

摘　　要：目的应用RNA干扰技术(RNAi)研究针对CD105的小片段RNA(siRNA)抑制CD105在体外培养的人脐静脉内皮细胞(HUVECs)中表达的情况,观察CD105沉默后对细胞生长的影响。方法根据人的CD105mRNA序列选择3个不同的靶位点,分别合成3条CD105siRNA,并同时合成阴性对照和阳性对照siRNA。采用脂质体介导的基因法转染至体外培养的HUVECs中,采用RT-PCR检测各组细胞CD105表达水平,Western blot检测干扰前后CD105蛋白表达水平,并用MTT法检测人HUVECs的细胞活力。结果 RT-PCR和Western blot检测表明,3号CD105siRNA能最有效沉默CD105的表达,转染后的HUVECs中CD105mRNA水平和蛋白表达水平明显下降。MTT结果表明CD105的下调导致了HUVECs活力的下降。结论特异性的siRNA可以下调HUVECs中CD105mRNA及蛋白表达水平,减低细胞活力。也提示CD105可能对体外培养HUVECs的生长有促进作用,CD105可以作为治疗新生血管的靶点。Objective To investigate the effect of silencing CD105 gene expression with siRNA on the proliferation of human umbilical vein endothelial cells（HUVECs）and CD105 mRNA and protein expression.Methods HUVECs were cultured in vitro and transfected with siRNA of CD105 by lipofectamine 2000.The expression levels of CD105 mRNA and protein before and after transfection were detected by semi-quantitative reverse transcription polymerase chain reaction（RT-PCR）and Western blot respectively.Apoptosis of HUVECs induced by CD105 siRNA was assayed by MTT.Results The expression level of CD105 mRNA in HUVECs was inhibited by the special siRNA.The expression of CD105 protein was down-regulated in HUVECs after interfered with siRNA.The cell proliferation in RNAi group was significantly lower than that in the negative control and control groups.Conclusion RNA interference can down-regulate the level of CD105 mRNA and protein expression,inducing the apoptosis of HUVECs.CD105 can accelerate the growth of HUVECs.CD105 may be used as a gene therapy target on corneal neovascularization.

关 键 词：RNA干扰 CD105基因 人脐静脉内皮细胞 基因沉默 

分 类 号：R349.64[医药卫生—基础医学]