Inhibition of NF-kappa B-Rel A expression by antisense oligodeoxynucleotides suppresses synthesis of urokinase-type plasminogen activator (uPA) but not its inhibitor PAI-1.
Inhibition of NF-kappa B-Rel A expression by antisense oligodeoxynucleotides suppresses synthesis of urokinase-type plasminogen activator (uPA) but not its inhibitor PAI-1.
复制标题
反义寡脱氧核苷酸对 NF-κ B-Rel A 表达的抑制会抑制尿激酶型纤溶酶原激活剂 (uPA) 的合成，但不会抑制其抑制剂 PAI-1。
作者：
U. Reuning;O. Wilhelm;T. Nishiguchi;L. Guerrini;F. Blasi;H. Graeff;M. Schmitt
尿激酶型纤溶酶原激活剂（uPA）在肿瘤侵袭和转移中的重要作用强调了对其表达进行微调细胞控制的必要性。研究表明，uPA 的变化与细胞侵袭力的变化直接相关。我们通过使用反义 (AS) 寡脱氧核苷酸 (ODN) 技术抑制 Rel 相关蛋白的表达，研究了 Rel 相关蛋白在人卵巢癌细胞 uPA 合成中的作用。将 OV-MZ-6 细胞暴露于针对 Rel A 的 10 microM 硫代磷酸 (PS) 衍生的 AS-ODN 中，导致细胞裂解物中 uPA 抗原最大减少 50%，细胞培养物上清液减少 70%，同时 uPA mRNA 水平显着短暂下降。针对 NF-kappa B1 (p50) 或 c-rel 的反义-PS-ODN 对 uPA 蛋白表达没有影响。针对 Rel A 的 AS-PS-ODN 也影响 OV-MZ-6 细胞的蛋白水解能力，反映为与未处理的对照相比，24 小时内细胞的纤维蛋白溶解能力下降约 70%。 AS-PS-ODN 定向 I kappa B α 表达，使细胞培养物上清液中的 uPA 增加高达 50%。应用任一 AS-PS-ODN 均不会改变 uPA 受体 (uPAR) 的产生和 1 型纤溶酶原激活剂抑制剂 (PAI-1) 的合成。蛋白质印迹和凝胶延迟分析揭示了 OV-MZ-6 细胞核蛋白提取物中 Rel 相关蛋白的组成型表达。因此，这些蛋白质似乎与 uPA 调节有关，从而可能有助于肿瘤扩散和转移。
The essential role of urokinase-type plasminogen activator (uPA) in tumor invasion and metastasis stresses the necessity of a fine-tuned cellular control over its expression. It has been shown that changes in uPA directly correlate with changes in cell invasiveness. We examined the role of Rel-related proteins in uPA synthesis by human ovarian cancer cells by inhibiting their expression using the antisense (AS) oligodeoxynucleotide (ODN) technology. Exposure of OV-MZ-6 cells to 10 microM phosphorothioate (PS)-derivatized AS-ODN directed to Rel A led to a maximal 50% decrease of uPA antigen in cell lysates and a 70% reduction in cell cultures supernatants accompanied by a significant transient decline in uPA mRNA levels. Antisense-PS-ODN directed to NF-kappa B1 (p50) or c-rel had no effect on uPA protein expression. AS-PS-ODN directed to Rel A also affected the proteolytic capacity of OV-MZ-6 cells reflected by an approximately 70% decrease in the fibrinolytic capacity of the cells within 24 h compared to untreated controls. AS-PS-ODN directed to I kappa B alpha expression increased uPA in cell culture supernatants up to 50%. uPA receptor (uPAR) production and synthesis of plasminogen activator inhibitor type-1 (PAI-1) were not altered by either AS-PS-ODN applied. Western blot and gel retardation analyses revealed constitutive expression of Rel-related proteins in nuclear protein extracts of OV-MZ-6 cells. Thus these proteins seem to be implicated in uPA regulation and may thereby contribute to tumor spread and metastasis.