FOXQ1介导TGF-β1信号通路调控胰腺癌PANC-1细胞体外血管生成  

FOXQ1 gene mediates TGF-β1 signaling pathway to regulate in vitro angiopoiesis of pancreatic cancer PANC-1 cells

作　　者：邓大炜 吴斌 严舒[1,2] 兰川 张光年[1,2] 弋鹏圣 曾丽娟[3] 李建水 

机构地区：[1]川北医学院肝胆胰肠研究所,四川南充637000 [2]川北医学院附属医院肝胆外科,四川南充637000 [3]川北医学院信息科,四川南充637000

出　　处：《中国肿瘤生物治疗杂志》2018年第1期68-72,共5页Chinese Journal of Cancer Biotherapy

基　　金：四川省教育厅科研项目资助(No.12ZB218)~~

摘　　要：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-sh RNA重组慢病毒和阴性对照慢病毒(NC-sh RNA)感染PANC-1细胞,流式细胞术检测感染率,实时荧光定量PCR(q PCR)和Western blotting法检测沉默效果。实验设FOXQ1-sh RNA组、NC-sh RNA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q PCR、Western blotting法检测VEGF-A、MMP-2 m RNA和蛋白的表达水平;用TGF-β1(终质量浓度5 ng/ml)诱导FOXQ1-sh RNA组和NC-sh RNA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:sh RNA慢病毒感染率为90%左右,FOXQ1-sh RNA组PANC-1细胞的体外血管生成数目显著少于NC-sh RNA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 m RNA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 Q 1 （FOXQ1） gene silencing on in vitro angiopoiesis of pancreatic carcino- ma cells, and to investigate its role in transforming growth factor-β1（TGF-β1） signaling pathway. Methods： FOXQI-shRNA recombi- nant 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. FOXQI-shRNA group, NC-shRNA group and blank group were set up. Human umbilical vein endothelial cells （HUVECs） were used for the in vitro angiopoie- sis assay, which was observed under fluorescence microscopy. Meanwhile, qPCR and Western blotting were performed to examine mRNA and protein expressions of FEGF-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 efIiciency of lentivirus was about 90%. Compared with NC-shRNA group, the angiopoiesis ability in FOXQI-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 ofFOXQ1, FEGF-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 mecha- nism may be related to the down-regulation of VEGF-A and MMP-2 and possibly be regulated by TGF-β1 pathway.

关 键 词：胰腺癌 PANC-1细胞 叉头框蛋白Q1基因 转化生长因子-β1(TGF-β1) 血管生成 

分 类 号：R735.9[医药卫生—肿瘤] R730.2[医药卫生—临床医学]