[1]许剑锋,沈维干,彭琬昕,等.腺病毒介导的RNA干扰降低VEGF转录抑制血管形成的研究[J].南京师范大学学报(工程技术版),2007,07(04):084-87.
　Xu Jianfeng,Shen Weigan,Peng Wanxin,et al.Study on Inhibitation of the Tube Formation by RNA Introduced by Adenovirus by its Interference With and Decrease of VEGF Transcription[J].Journal of Nanjing Normal University(Engineering and Technology),2007,07(04):084-87.
点击复制

腺病毒介导的RNA干扰降低VEGF转录抑制血管形成的研究
分享到：

南京师范大学学报（工程技术版）[ISSN:1006-6977/CN:61-1281/TN]

卷:
07卷
期数:
2007年04期
页码:
084-87
栏目:
出版日期:
2007-12-30

文章信息/Info

Title:
Study on Inhibitation of the Tube Formation by RNA Introduced by Adenovirus by its Interference With and Decrease of VEGF Transcription
作者:
许剑锋;沈维干;彭琬昕;李朝军;
南京师范大学生命科学学院, 江苏南京210097
Author(s):
Xu Jianfeng; Shen Weigan; Peng Wanxin; Li Chaojun
School of Life Science,Nanjing Normal University,Nanjing 210097,China
关键词:
血管生成;  血管内皮生长因子;  RNA干扰
Keywords:
ang iogenesis;  VEGF;  RNA i
分类号:
Q789
摘要:
用细菌内同源重组的方法构建了针对VEGF的siRNA表达重组腺病毒AdH1-siRNA/VEGF,感染HUVEC细胞,观察该病毒对HUVEC细胞体外增殖和形成微血管的干扰作用.通过Real-timeRT-PCR实验表明,AdH1-siRNA/VEGF可特异性地下调血管内皮细胞(HUVEC)中的VEGFmRNA水平.与对照组相比,VEGF的mRNA水平下降为50%,AdH1-siRNA/VEGF对HUVEC细胞增殖有干扰作用.加入病毒9d后,对照组的细胞平均计数为31.5×104/μL,干扰病毒对照组为28×104/μL,AdH1-siRNA/VEGF干扰组为21.5×104/μL.AdH1-siRNA/VEGF对HUVEC细胞在Matrigel上形成微血管有干扰作用,对照组每HPF形成微血管数量为10.75条,干扰病毒对照组为10.25条,实验组AdH1-siRNA/VEGF每HPF形成微血管数量为7条.证明AdH1-siRNA/VEGF可以干扰血管形成.
Abstract:
This study uses the hom o logous recomb ination technique to construct a recomb inan t adenov irusAdH 1-siRNA /VEGF ta rgeting VEGF infects hum an endo the lia l ce lls ( HUVEC ), obse rves the inter fe rence o f the adenov irus w ith the pro liferation ofHUVEC in v itro and the tube fo rm ation. The exper iem ent of rea-l tim e reve rse transcr iption- PCR show s tha t this adenov irus can espec ia lly decrease level o f the VEGF mRNA o fHUVEC. Com pared w ith control g roup, themRNA expression o fVEGF decreased to 50%. AdH 1-siRNA /VEGF can effic iently inte rfere the pro lifeya-t ion o fHUVEC. N ine day s after infection o f adenov irus, the ave rage cell number o f contro l g roup and vector g roup are 31. 5 × 104 /LL and 28 × 104 /LL respective ly, that o f groups respective ly infected w ith AdH 1-siRNA /VEGF are 21. 5 × 104 /LL, and can effic iently inte rfere the tube form ation o fHUVEC onM aty ig e.l Ave rage ly, the tube number of everyH PF is 10. 75 in the contro l g roup, and 10. 25 in the vecto r group. In the group treated w ith AdH 1-siRNA / VEGF, there are 7 tubes in theHPE. AdH 1-siRNA /VEGF can inte rfere tube form a tion.

参考文献/References:

[ 1] Fo lkman J. Ang iogenes is in cancer, vascular, rheum a to id and o ther disease[ J]. NatM ed, 1995, 1( 1): 27- 31.
[ 2] Fe rrara N, H enze lW J. P itu itary fo llicular ce lls secre te a nove l hepar in-b inding grow th factor spec ific for vascular endothe lial ce lls[ J]. B iochem B iophys Res Commun, 1989, 161( 2): 851- 858.
[ 3] H annon G J. RNA interference[ J]. Nature, 2002, 418( 6894): 244- 251.
[ 4] Shen C, Buck A K, Liu X, et a .l Gene silenc ing by adenov irus-de livered siRNA[ J]. FEBS Lett, 2003, 539: 111- 114.
[ 5] Sam brook J, Fritsch E F, M aniatis T. Mo lecu la r C loning[M ]. 2nd ed. Co ld Spr ing H arbor, New Yo rk: Co ld Spring H arbor Labo ra tory Press, 1989.
[ 6] Annabi B, Thibeault S, Lee Y T, et a.l M atr ix m e tallopro te inase regulation of sph ingosine-1-phosphate- induced ang iogenic properties o f bone m arrow strom al ce lls[ J] . Exp H em ato,l 2003, 31: 640- 649.
[ 7] Zhang L, Yang N, M oham ed-H adleyA, et a.l Vecto r-based RNA,i a nove l tool fo r iso fom-specific kno ck-down of VEGF and ant-i ang iogenes is g ene therapy of cance r[ J]. B iochem B iophys Res Commun, 2003, 303: 1 169- 1 178.
[ 8] Re ich S J, Fo sno t J, Kurok iA, et a.l Sm a ll inter fe ring RNA ( siRNA) targetingVEGF effectively inhabits o cu lar neovasculariza tion in a m ousem oda l[ J] . M o lV is, 2003, 9: 210- 216.

备注/Memo

备注/Memo:
作者简介: 许剑锋( 1964-) , 硕士, 主要从事分子生物学方面的学习与研究. E-m a il: xjf646566@ sina. com
通讯联系人: 李朝军( 1966-) , 研究员, 博士生导师, 主要从事细胞生物学方面的教学与研究. E-ma il:C haojun Le@ Yah oo. com
更新日期/Last Update: 2013-04-29