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[诱饵寡脱氧核苷酸对NIH3T3细胞中胶原蛋白基因表达抑制作用的实验研究]

[An experimental study of inhibiting effect of decoy oligodeoxynucleotides on the gene expression of collagen in NIH3T3 cell].

作者信息

Hao Tian-zhi, Liang Huan-ping, Lv Feng-lin, Xu Xiang, Guo Hua, Wang Fu-long, Yang Wen-jun, Luo Yan, Li Lei

机构信息

Department of neurosurgery, Research Institute of surgery, Daping Hospital, Third military Medical University, Chongqing 400042, PR China.

出版信息

Zhonghua Shao Shang Za Zhi. 2003 Jun;19(3):175-8.

Abstract

OBJECTIVE

To investigate the effect of activator protein-1 (AP-1) decoy-oligodeoxynucleotides (Decoy-ODNs) on the expression of fibroblast alpha2 type I collagen, so as to explore the gene therapy of pathologic scar.

METHODS

Decoy-ODNs targeting AP-1 were designed and synthesized. NIH3T3 cells were transfected by cationic liposomes. The distribution of Decoy-ODNs in the cells was investigated. The inhibiting effects of Decoy-ODNs on AP-1 were determined by electrophoretic mobility shift assay (EMSA). And the effects of Decoy-ODNs on the collagen synthesis in the cells were analyzed by RT-PCR.

RESULTS

AP-1 Decoy-ODNs could competitively inhibit the AP-1 in vitro activity. Cationic liposomes could play roles by effectively transfecting Decoy-ODNs into the plasma and nucleus. The mRNA expression of fibroblast alpha2 type I collagen decreased evidently after 24 hours of Decoy-ODNs action.

CONCLUSION

Decoy-ODNs could inhibit the mRNA expression of fibroblast alpha2 type I collagen by antagonizing AP-1.

摘要

目的

研究活化蛋白-1(AP-1)诱饵寡脱氧核苷酸(Decoy-ODNs)对成纤维细胞α2Ⅰ型胶原表达的影响,以探讨病理性瘢痕的基因治疗方法。

方法

设计并合成针对AP-1的Decoy-ODNs。采用阳离子脂质体转染NIH3T3细胞,研究Decoy-ODNs在细胞内的分布情况。通过电泳迁移率变动分析(EMSA)检测Decoy-ODNs对AP-1的抑制作用。运用逆转录聚合酶链反应(RT-PCR)分析Decoy-ODNs对细胞内胶原合成的影响。

结果

AP-1 Decoy-ODNs能够在体外竞争性抑制AP-1的活性。阳离子脂质体可通过将Decoy-ODNs有效转染至细胞质及细胞核发挥作用。Decoy-ODNs作用24小时后,成纤维细胞α2Ⅰ型胶原的mRNA表达明显降低。

结论

Decoy-ODNs可通过拮抗AP-1抑制成纤维细胞α2Ⅰ型胶原的mRNA表达。

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