ZNF580过表达对内皮细胞血管内皮生长因子的表达及增殖能力的影响

【目的】研究人锌指蛋白新基因ZNF580过表达对人脐静脉内皮细胞株EA.hy926血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达及增殖能力的变化,为探讨ZNF580基因功能提供科学依据。【方法】重组质粒pEGFP-ZNF580经LipofectamineTM2000脂质体转染法转染EA.hy926细胞,48h后荧光显微镜下观察ZNF580-EGFP在内皮细胞中的表达情况;应用半定量RT-PCR、Western blotting及ELISE法检测转染前后内皮细胞ZNF580和VEGFmRNA、蛋白表达的变化;运用MTT实验检测转染前后内皮细胞增殖能力的改变。【结果】荧光显微镜下示融合蛋白ZNF580-EGFP在EA.hy926细胞核中表达;ZNF580基因过表达后,内皮细胞VEGFmRNA及蛋白的表达均上调(P<0.05),且其增殖能力显著升高(P<0.01)。【结论】ZNF580可以调控EA.hy926细胞VEGF的表达,且ZNF580在内皮细胞增殖过程中具有重要的调节作用。

Effects of ZNF580 gene over expression on endothelial cells

【Objective】To observe the effects of the novel human C2H2-zinc finger protein gene ZNF580 overexpression on the expression of VEGF and the changes of proliferation of endothelial cells.【Methods】The plasmids pEGFP-ZNF580 was transfected into EA.hy926 cells with Lipofectamine 2000 for 48 h. The localization was estimated by fluorescence microscope. The mRNA and protein expression of ZNF580 and VEGF before and after transfection were detected by RT-PCR, Western blot and ELISE respectively. The viability of EA.hy926 cells was determined by MTT method.【Results】The fusion protein of EGFP-ZNF580 was localized in the nuclei of EA.hy926cells. The VEGF mRNA and protein levels were both elevated in EAhy926 cells of overexpression of ZNF580 by RT-PCR analysis and ELISA（P 0.05）. Compared to EAhy926 cells, EA.hy926 cells transiently transfected with pEGFP-ZNF580 exhibited a significant increase in the proliferation （P 0.01）.【Conclusion】ZNF580 could regulate the expression of VEGF, furthermore ZNF580 might play an important role in the process of the proliferation of endothelial cells.