摘要
【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.
出处
《武警医学院学报》
CAS
2011年第7期521-524,F0003,共5页
Acta Academiae Medicinae CPAPF
基金
国家自然科学基金资助项目(30770885)
天津市应用基础及前沿技术研究计划项目(10JCYBJC26500)
武警医学院面上科研项目(WY200903)