Park Kwan-Kyu, Deok Ahn Jong, Lee In-Kyu, Magae Junji, Heintz Nicholas H, Kwak Jong-Young, Lee Young-Choon, Cho Young-Su, Kim Hyun-Chul, Chae Young-Mi, Ho Kim Young, Kim Cheorl-Ho, Chang Young-Chae
Kidney Institute, Keimyung University School of Medicine, 194, Dongsan-Dong, Jung-Gu, Daegu 700-712, South Korea.
Biochem Biophys Res Commun. 2003 Sep 5;308(4):689-97. doi: 10.1016/s0006-291x(03)01455-4.
Proliferation of glomerular mesangial cells (MCs) is an important feature of several forms of glomerulonephritis. The transcription factor E2F coordinately regulates expression of genes required for cell proliferation, thereby mediating cell growth control. Here we investigated the role of E2F1 and E2F4 expression, with or without co-expression of DP1 or DP2, on cell proliferation in transiently transfected primary rat MCs. In transfected cells, cell proliferation induced by over-expression of E2F was significantly enhanced by co-expression of DP proteins. Previous studies showed that the transfection of decoy oligodeoxynucleotides (ODNs) corresponding to E2F binding sites inhibits cell proliferation. Here we have developed a Ring-E2F (R-E2F) decoy ODN with a circular dumbbell structure and compared its effects with those of a phosphorothioated E2F decoy (PS-E2F decoy) ODN. The R-E2F decoy ODN showed enhanced stability in the presence of nucleases and sera, and inhibited E2F/DP-dependent promoter activity of cell cycle genes more effectively than the PS-E2F decoy ODN. Transfection of R-E2F decoy ODN resulted in strong inhibition of cell cycle gene expression and MC proliferation. Our data suggest that E2F/DP complexes play a critical role in the MC proliferation and that the R-E2F decoy ODN may be a powerful tool for inhibiting cell proliferation.
肾小球系膜细胞（MCs）增殖是多种形式肾小球肾炎的一个重要特征。转录因子E2F协同调节细胞增殖所需基因的表达，从而介导细胞生长控制。在此，我们研究了E2F1和E2F4表达（有无DP1或DP2共表达）对瞬时转染的原代大鼠MCs细胞增殖的作用。在转染细胞中，DP蛋白共表达显著增强了E2F过表达诱导的细胞增殖。先前的研究表明，转染与E2F结合位点对应的诱饵寡脱氧核苷酸（ODN）可抑制细胞增殖。在此，我们开发了一种具有环状哑铃结构的环状E2F（R-E2F）诱饵ODN，并将其效果与硫代磷酸化E2F诱饵（PS-E2F诱饵）ODN进行比较。R-E2F诱饵ODN在核酸酶和血清存在下显示出增强的稳定性，并且比PS-E2F诱饵ODN更有效地抑制细胞周期基因的E2F/DP依赖性启动子活性。转染R-E2F诱饵ODN导致细胞周期基因表达和MC增殖受到强烈抑制。我们的数据表明，E2F/DP复合物在MC增殖中起关键作用，并且R-E2F诱饵ODN可能是抑制细胞增殖的有力工具。
