Maeshima Y, Kashihara N, Yasuda T, Sugiyama H, Sekikawa T, Okamoto K, Kanao K, Watanabe Y, Kanwar Y S, Makino H
Department of Medicine III, Okayama University Medical School, Okayama 700, Japan.
J Clin Invest. 1998 Jun 1;101(11):2589-97. doi: 10.1172/JCI429.
The transcription factor E2F coordinately activates several cell cycle-regulatory genes. We attempted to inhibit the proliferation of mesangial cells in vitro and in vivo by inhibiting E2F activity using a 25-bp decoy oligodeoxynucleotide that contained consensus E2F binding site sequence (E2F-decoy) as a competitive inhibitor. The decoy's effect on human mesangial cell proliferation was evaluated by [3H]thymidine incorporation. The E2F decoy inhibited proliferation in a concentration-dependent manner, whereas a mismatch control oligodeoxynucleotide had little effect. Electrophoretic mobility shift assays demonstrated that the decoy's inhibitory effect was due to the binding of the decoy oligodeoxynucleotide to E2F. The effect of the E2F decoy was then tested in a rat anti-Thy 1.1 glomerulonephritis model. The E2F decoy oligodeoxynucleotide was introduced into the left kidney 36 h after the induction of glomerulonephritis. The administration of E2F decoy suppressed the proliferation of mesangial cells by 71%. Furthermore, treatment with the E2F decoy inhibited the glomerular expression of proliferating cell nuclear antigen at the protein level as well as the mRNA level. These findings indicate that decoy oligonucleotides can suppress the activity of the transcription factor E2F, and may thus have a potential in treating glomerulonephritis.
转录因子E2F协同激活多个细胞周期调控基因。我们试图通过使用一种含有E2F结合位点共有序列的25个碱基对的诱饵寡脱氧核苷酸（E2F诱饵）作为竞争性抑制剂来抑制E2F活性，从而在体外和体内抑制系膜细胞的增殖。通过[3H]胸苷掺入法评估诱饵对人系膜细胞增殖的影响。E2F诱饵以浓度依赖的方式抑制增殖，而错配对照寡脱氧核苷酸几乎没有作用。电泳迁移率变动分析表明，诱饵的抑制作用是由于诱饵寡脱氧核苷酸与E2F结合所致。然后在大鼠抗Thy 1.1肾小球肾炎模型中测试E2F诱饵的作用。在诱导肾小球肾炎36小时后，将E2F诱饵寡脱氧核苷酸导入左肾。给予E2F诱饵可使系膜细胞增殖抑制71%。此外，用E2F诱饵处理在蛋白质水平和mRNA水平上均抑制了增殖细胞核抗原的肾小球表达。这些发现表明，诱饵寡核苷酸可以抑制转录因子E2F的活性，因此可能在治疗肾小球肾炎方面具有潜力。
