siRNA沉默ZNF580基因对内皮细胞MMP-2的表达及侵袭能力的影响

Effects of ZNF580 gene silenced by small interfering RNA on endothelial cells

【目的】观察ZNF580基因沉默后内皮细胞MMP-2的表达及迁移和侵袭能力的变化,为探讨ZNF580基因功能提供科学依据。【方法】针对ZNF580基因不同部位设计并化学合成3对不同靶向小干扰RNA(siRNA),脂质体介导瞬时转染EA.hy926内皮细胞;应用半定量RT-PCR、Western blotting及明胶酶谱检测转染前后内皮细胞ZNF580和MMP-2 mRNA、蛋白的表达及活性变化;运用划痕愈合实验和Transwell小室模型检测转染前后内皮细胞迁移和侵袭能力的改变。【结果】筛选出最佳干扰序列siRNA序列3;下调ZNF580基因表达后,MMP-2 mRNA的表达及明胶酶的活性均下调(P<0.05),且内皮细胞体外迁移和侵袭能力显著降低(P<0.01)。【结论】ZNF580在内皮细胞运动转移过程中具有重要作用,可能成为肿瘤基因治疗的一个新靶点。

[Objective]To observe the effects of ZNF580 gene silenced by small interfering RNA（siRNA） on the expression of MMP-2 and the changes of migrative and invasive abilities of endothelial cells. [ Methods ] EA.hy926 endothelial cells were transfected in vitro with sequence-specific siRNA targeting ZNF580 gene using RNA interference technique. The mRNA and protein expression of ZNF580 and MMP-2 before and after transfection were detected by RT-PCR,Western blot and gelatin zymography respectively. The migration and invasion abilities of cells were detected by wound healing assay and transwell assay. [Results] The expressions of ZNF580 and MMP-2 were both decreased（P〈0.05）. The migrative and invasive abilities were reduced obviously （P〈0.01）. [ Conclusion ]ZNF580 might play an important role in endothelial cells migration and invasion, which indicates that ZNF580 gene may be considered as one of the target for gene therapy of cancer.