Ang2、Tie2基因的RNA干扰及其在体外抑制血管生成的作用  

Ang2,Tie2 RNA Interference and the Function of Them on Inhibiting Angiogenesis in vitro

作　　者：王彪[1] 刘照亮[1] 林菊丽[1] 单秀英[1] 王美水[1] 张声[1] 鲁开化[2] 

机构地区：[1]福建医科大学附属第一医院,福州350005 [2]第四军医大学西京医院整形外科研究所,西安710043

出　　处：《中国生物工程杂志》2010年第4期20-25,共6页China Biotechnology

基　　金：卫生部科学研究基金-福建省卫生教育联合攻关计划(WKJ2008-2-51);福建省自然科学基金计划(2007J0269);福建省教育厅科技项目(JB08088)资助项目

摘　　要：目的:探讨应用RNA干扰(RNA interference,RNAi)技术沉默Ang2、Tie2基因及其在体外抑制血管生成的研究,为将来进一步进行抑制肿瘤血管生成的动物实验研究奠定基础,为肿瘤的基因治疗提供实验依据。方法:用pSilencer1.0-U6-Ang2/Tie2-siRNA重组质粒转染人脐静脉内皮细胞(HUVECs),采用RT-PCR检测各组HUVECsAng2、Tie2的mRNA表达状况。应用体外血管生成的三维培养模型研究转染后的HUVECs在体外形成血管样结构的情况。结果:pSilencer1.0-U6-Ang2/Tie2-siRNA重组质粒转染HUVECs后,RT-PCR检测结果显示:Ang2、Tie2基因mRNA的表达水平均受到明显抑制(P<0.05),并且siRNA的2条重组质粒之间均无明显差别(P>0.05)。转染后的HUVECs在体外三维培养模型中血管样结构形成的数量和长度均明显减少(P<0.05),表明血管生成受到明显抑制。结论:Ang2-siRNA、Tie2-siRNA能够抑制HUVECs中Ang2和Tie2的mRNA表达,从而在体外抑制血管生成。Objective：The expression of Ang2 and Tie2 is silenced by RNA interference in order to inhibit the angiogenesis in vitro,this will supply the theory basis to the further animal research on the inhibition of tumor angiogenesis in vivo and provide the experimental evidence for tumor gene therapy in future.Methods：Using pSilencer 1.0-U6-Ang2/Tie2-siRNA recombinant plasmid to transfect into human umbilical vein endothelial cells （HUVECs）,using RT-PCR to analyze the mRNA expression levels of Ang2 and Tie2 in HUVECs of each groups respectively.And the formation of vascular-like structure in HUVECs,transfected by the plasmids,was studied in vitro by three-dimensional culture.Results：After the recombinant plasmids of pSilencer1.0-U6-Ang2,Tie2-siRNA transfected HUVECs,the mRNA expression levels of Ang2,Tie2 in HUVECs were average obviously suppressed by RT-PCR（P0.05）,and there is no significant difference on the expression levels（P0.05）between the two plasmids of each siRNA.There is a significant decrease on the quantity and length of vascular-like structure in HUVECs,transfected with the recombinant plasmids by three-dimensional culture model in vitro,（P0.05）.It proves that angiogenesis has been inhibited significantly in vitro.Conclusions：Ang2-siRNA,Tie2-siRNA can inhibit the mRNA expression of Ang2,Tie2 in HUVECs,and inhibit angiogenesis in the model of three-dimensional culture in vitro.

关 键 词：RNAI 促血管生成素家族及其受体 血管生成 内皮细胞体外三维培养 

分 类 号：Q819[生物学—生物工程]