An oligonucleotide decoy for transcription factor E2F inhibits mesangial cell proliferation in vitro.
An oligonucleotide decoy for transcription factor E2F inhibits mesangial cell proliferation in vitro.
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转录因子 E2F 的寡核苷酸诱饵可在体外抑制系膜细胞增殖。
DOI：
10.1152/ajprenal.1998.275.2.f278
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Dzau,VJ
中科院分区：
文献类型：
--
作者：
Tomita,N;Horiuchi,M;Tomita,S;Gibbons,GH;Kim,JY;Baran,D;Dzau,VJ
转录因子E2F控制几个与细胞增殖有关的基因的表达，包括c-myc、c-myb、增殖细胞核抗原（PCNA）和cdk2激酶。在确定PCNA和cdk2激酶在大鼠系膜细胞（MC）中通过血清刺激诱导后，我们试图通过用含有合成的双链寡脱氧核苷酸（ODN）的阳离子脂质体转染这些细胞来抑制MC的体外增殖，该脂质体与E2F具有高亲和力。通过凝胶迁移转移试验，我们检测到血清刺激后MC中E2F的特异性结合增加。MC核提取物与E2F高亲和力的双链ODN预孵育可完全抑制这种结合，但与含有两点突变的错义ODN预孵育不能完全抑制这种结合。MC也被含有三个E2F结合位点的荧光素酶报告基因构建体转染。血清刺激MC可增强荧光素酶活性，而与E2F诱饵ODN共转染MC可特异性消除这种作用。此外，RT-PCR分析显示，转染E2F诱饵ODN可抑制血清诱导的PCNA和cdk2激酶基因表达上调，而转染错义ODN则无抑制作用。这些基因表达的变化与E2F诱饵ODN转染细胞中PCNA和cdk2激酶蛋白表达的减少相一致。血清刺激后MC数增加。转染E2F诱饵ODN可以减弱这种作用，而转染错义ODN则不能。这些数据表明转录因子E2F在MC增殖调控中起着至关重要的作用，并且该因子可以成功靶向抑制MC细胞周期进程。
The transcription factor E2F controls expression of several genes involved in cell proliferation including c-myc, c-myb, proliferating cell nuclear antigen (PCNA), and cdk2 kinase. Having established that both PCNA and cdk2 kinase are induced in rat mesangial cells (MC) by serum stimulation, we attempted to inhibit MC proliferation in vitro by transfecting these cells with cationic liposomes containing a synthetic double-stranded oligodeoxynucleotide (ODN) with high affinity for E2F. Using a gel mobility shift assay, we detected increased specific binding of E2F in MC following serum stimulation. This binding was completely inhibited by preincubation of MC nuclear extracts with the double-stranded ODN with high affinity for E2F but not by preincubation with a missense ODN containing two point mutations. MC were also transfected with a luciferase reporter gene construct containing three E2F binding sites. Luciferase activity was enhanced by serum stimulation of MC, and this effect was specifically abolished by cotransfection of MC with E2F decoy ODN. Furthermore, RT-PCR analysis revealed that serum-induced upregulation of PCNA and cdk2 kinase gene expression was inhibited by E2F decoy ODN transfection but not by transfection of missense ODN. These changes in gene expression were paralleled by a reduction in PCNA and cdk2 kinase protein expression in E2F decoy ODN transfected cells. MC number increased following serum stimulation. This effect was blunted by transfection with E2F decoy ODN but not by transfection of missense ODN. These data suggest that the transcription factor E2F plays a crucial role in the regulation of MC proliferation and that this factor can be successfully targeted to inhibit MC cell cycle progression.
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DOI：
10.1073/pnas.86.10.3594
发表时间：
1989-05-01
影响因子：
11.1
作者：
HIEBERT, SW;LIPP, M;NEVINS, JR
通讯作者：
NEVINS, JR
cAMP 组织特异性调节小鼠肾素基因表达的分子机制。
DOI：
--
发表时间：
1991
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Horiuchi,M;Nakamura,N;Tang,SS;Barrett,G;Dzau,VJ
通讯作者：
Dzau,VJ
影响因子：
64.8
作者：
WEINTRAUB, SJ;PRATER, CA;DEAN, DC
通讯作者：
DEAN, DC
DOI：
10.1111/j.1349-7006.1992.tb00133.x
发表时间：
1992-06
期刊：
Japanese journal of cancer research : Gann
影响因子：
--
作者：
Yamaguchi M;Hayashi Y;Hirose F;Matsuoka S;Shiroki K;Matsukage A
通讯作者：
Matsukage A
影响因子：
15.9
作者：
ITOH, H;PRATT, RE;DZAU, VJ
通讯作者：
DZAU, VJ