目的：探讨沉默叉头框蛋白Q1（forkhead box Q1，FOXQ1）基因对胰腺癌PANC-1 细胞体外血管生成的影响及其在转化生长因子-β1（transforming growth factor-β1，TGF-β1）信号通路中的作用。方法：用FOXQ1-shRNA重组慢病毒和阴性对照慢病毒（NC-shRNA）感染PANC-1 细胞，流式细胞术检测感染率，实时荧光定量PCR（qPCR）和Western blotting 法检测沉默效果。实验设FOXQ1-shRNA组、NC-shRNA组与空白对照组，用人脐静脉内皮细胞（human umbilical vein endothelial cell，HUVEC）进行体外血管生成实验，荧光显微镜下观察细胞血管生成能力。用qPCR、Western blotting 法检测VEGF-A、MMP-2 mRNA和蛋白的表达水平；用TGF-β1（终质量浓度5 ng/ml）诱导FOXQ1-shRNA组和NC-shRNA组细胞，检测诱导前后细胞体外血管生成能力变化与FOXQ1、VEGF-A、MMP-2 表达差异。结果：shRNA慢病毒感染率为90%左右，FOXQ1-shRNA组PANC-1 细胞的体外血管生成数目显著少于NC-shRNA组[ （9.33±2.08）vs（28.67±2.52）条，P<0.05]，其VEGF-A、MMP-2 表达下调（均P<0.05）。TGF-β1 增强各组细胞体外血管生成能力，促进FOXQ1、VEGF-A、MMP-2 mRNA和蛋白的表达水平（均P<0.05）。结论：FOXQ1 介导了胰腺癌PANC-1 细胞体外血管生成，其机制可能与VEGF-A和MMP-2 的下调有关，且可能受TGF-β1 通路的调控。

Objective: To explore the effect of forkhead box Q1（FOXQ1）gene silencing on in vitro angiopoiesis of pancreatic carcinoma cells, and to investigate its role in transforming growth factor-β1(TGF-β1) signaling pathway. Methods: FOXQ1-shRNA recombinant lentiviral vector and negative control lentiviral vector (NC-shRNA) were transfected into PANC-1 cells. Flow cytometry was used to detect the transfection efficiency. The gene silencing efficiency was measured by qPCR and Western blotting. FOXQ1-shRNA group,NC-shRNA group and blank group were set up. Human umbilical vein endothelial cells (HUVECs）were used for the in vitro angiopoiesis assay, which was observed under fluorescence microscopy. Meanwhile, qPCR and Western blotting were performed to examine mRNA and protein expressions of VEGF-A and MMP-2, respectively. After induction by TGF-β1 (final concentration of 5 ng/ml), the changes in angiopoiesis ability as well as the expression changes in FOXQ1，VEGF-A，MMP-2 in each group were detected. Results:The transfection efficiency of lentivirus was about 90%. Compared with NC-shRNA group, the angiopoiesis ability in FOXQ1-shRNA group was remarkably decreased（9.33±2.08 vs 28.67±2.52，P<0.05); Meanwhile, the expressions of VEGF-A and MMP-2 were all declined significantly (P<0.05). TGF-β1 improved the mRNA and protein expressions of FOXQ1，VEGF-A and MMP-2, and increased the in vitro angiopoiesis ability (P<0.05). Conclusion: FOXQ1 gene could mediate the in vitro angiopoiesis of PANC-1 cells; its mechanism may be related to the down-egulation of VEGF-A and MMP-2 and possibly be regulated by TGF-β1 pathway.

四川省教育厅科研项目资助( No.12ZB218）