中文摘要：

目的探讨小泛素相关修饰蛋白特异性蛋白酶1（SENP1）对脐静脉内皮细胞（HUVEC）增殖、迁移及凋亡的影响。方法用20个感染复数的p LKO.1-SENP1-小发夹RNA（shRNA）-绿色荧光蛋白（GFP）干涉慢病毒（SENP1干涉组）和p PIG-U6-随机序列（scramble）-GFP对照慢病毒（GFP-SC对照组）感染HUVEC。病毒感染HUVEC 48 h后,用流式细胞仪检测慢病毒的感染效率,实时荧光定量PCR（RT-PCR）和Western蛋白质印迹法检测HUVEC内SENP1基因和蛋白表达水平。用CCK8试剂盒检测细胞增殖能力,Transwell小室法检测细胞的迁移能力,流式细胞仪分析细胞凋亡率。结果 GFP-SC对照组和SENP1干涉组感染效率均〉97%。与GFP-SC对照组比较,SENP1干涉组HUVEC SENP1基因和蛋白表达水平显著降低（P〈0.01）。CCK8法测定结果表明,SENP1干涉组HUVEC增殖能力较GFP-SC对照组明显降低（P〈0.01）。SENP1干涉组细胞迁移过Transwell小室的细胞数量明显低于GFP-SC对照组（P〈0.01）。用低浓度血清（含2%胎牛血清）培养基诱导细胞凋亡24 h,SENP1干涉组细胞凋亡率即已升高到GFP-SC对照组的2倍左右（P〈0.05）;诱导凋亡至48 h,SENP1干涉组细胞凋亡率升高到对照组的3.2倍左右（P〈0.05）。结论 HUVEC内SENP1表达水平的降低不仅可以显著抑制细胞增殖和迁移能力,亦可促进细胞凋亡。

英文摘要：

OBJECTIVE To investigate the effect of small ubiquitin-related modifier-specific prote- ase 1 （SENP1） on proliferation, migration and apoptosis of human umbilical vein endothelial cells （HUVECs）. METHODS 20 multiplicity of infection （MOI） pPIG-U6-scramble-green fluorescent protein （GFP） lentivirus （GFP-SC group） and pLKO. 1-SENPI-shRNA-GFP interfering lentivirus （sh-SENP1 group） were used to infect HUVECs. Forty-eight hours after infection, flow cytometry was used to evalu- ate the infection efficiency while real-time PCR and Western blotting were used to detect the gene and protein expression of SENP1 in HUVECs. The effect of sh-SENP1 on migration, proliferation and apopto- sis was examined via Transwell, CCK8 assay and flow cytometry, respectively. RESULTS The infec- tion efficiency of HUVECs in GFP-SC group and sh-SENP1 group was above 97%. Compared with GFP-SC group, interfering SENP1 could significantly decrease the intracellular expression levels of SENP1 gene and protein of HUVECs （ P〈0.01 ）. CCK8 assay showed that the proliferation of sh-SENP1 group was significantly decreased compared with GFP-SC group （P〈0.01 ）. The cell number of HUVECs in sh-SENP1 group which migrated into the Transwell chamber was significantly smaller than that of GFP-SC group （ P〈0.01 ）. After a medium containing 2% fatal bovine serum was used to induce apoptosis for 24 h, the apoptosis rate of sh-SENP1 group was about 2 fold that of GFP-SC group （P〈0.05） and the difference reached 3.2 fold at 48 h （ P〈0.01 ）. CONCLUSION Inhibition of SENP1 can not only inhibit the migration and proliferation of HUVECs, but also promote the apoptosis of HUVECs.