Reuning U, Wilhelm O, Nishiguchi T, Guerrini L, Blasi F, Graeff H, Schmitt M
Frauenklinik, Technischen Universität München, Klinikum rechts der Isar, Germany.
Nucleic Acids Res. 1995 Oct 11;23(19):3887-93. doi: 10.1093/nar/23.19.3887.
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.
尿激酶型纤溶酶原激活剂（uPA）在肿瘤侵袭和转移中起着至关重要的作用，这凸显了对其表达进行精确细胞调控的必要性。研究表明，uPA的变化与细胞侵袭性的变化直接相关。我们通过使用反义（AS）寡脱氧核苷酸（ODN）技术抑制人卵巢癌细胞中Rel相关蛋白的表达，来研究它们在uPA合成中的作用。将OV-MZ-6细胞暴露于针对Rel A的10 microM硫代磷酸酯（PS）衍生的AS-ODN中，导致细胞裂解物中uPA抗原最大减少50%，细胞培养上清液中减少70%，同时uPA mRNA水平显著短暂下降。针对NF-κB1（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也影响了OV-MZ-6细胞的蛋白水解能力，与未处理的对照相比，24小时内细胞的纤溶能力下降了约70%。针对IκBα表达的AS-PS-ODN使细胞培养上清液中的uPA增加了50%。所应用的任何一种AS-PS-ODN都未改变uPA受体（uPAR）的产生以及1型纤溶酶原激活剂抑制剂（PAI-1）的合成。蛋白质印迹和凝胶阻滞分析显示，Rel相关蛋白在OV-MZ-6细胞核蛋白提取物中组成性表达。因此，这些蛋白似乎与uPA的调节有关，从而可能促进肿瘤的扩散和转移。
