FOXQ1基因介导了Shh诱导的SW480细胞血管生成及增殖

FOXQ1 gene mediate angiogenesis and proliferation induced by Shh in SW480 cells

目的:探讨FOXQ1基因沉默对大肠癌SW480细胞血管生成及增殖能力的影响及其在Sonic hedgehog(Shh)通路中的作用。方法:以携带FOXQ1-shRNA的慢病毒载体感染SW480细胞,将细胞株分为FOXQ1-shRNA和NC-shRNA组,利用脐静脉内皮细胞系926细胞行体外血管形成实验,荧光显微镜观察2组细胞血管形成能力,MTT、倍增时间、平板克隆形成实验检测2组细胞的存活率,real-time PCR、Western blot检测血管内皮生长因子(VEGF)-A、基质金属蛋白酶(MMP)2和cyclin D1 mRNA和蛋白水平。利用重组Shh蛋白诱导上述2组细胞,观察诱导前后细胞存活率及血管形成能力的变化,检测上述分子的表达差异。结果:与NC-shRNA组细胞相比,FOXQ1-shRNA组细胞的体外血管形成能力降低,而存活率无明显变化,real-time PCR及Western blot显示FOXQ1基因沉默后,VEGF-A和MMP2的表达下调,cyclin D1表达无显著差异。与NC-shRNA组比较,重组Shh蛋白诱导的FOXQ1-shRNA体外血管形成能力更低,存活率无明显差异。结论:FOXQ1基因介导了大肠癌SW480细胞的血管形成能力,对存活率无明显影响,并且可能受到Shh通路调控。

AIM：To explore the effect of FOXQ1 gene silencing on angiogenesis and proliferation ability of colon cancer cells induced by Sonic hedgehog（ Shh）.METHODS：Lentivirus expressing different FOXQ1-shRNA or negative cantrol（ NC）-shRNA was used to infect the SW480 cells.The best silencing condition was screened and used in the following experiments.The SW480 cells were divided into interfered group（ FOXQ1-shRNA） and control group（ NC-shRNA）.The MTT assay was used to observe the doubling time and cell activity.Tube formation assay was performed to detect the ability of angiogenesis.Meanwhile,the expression of vascular endothelial growth factor（ VEGF）-A,matrix metalloproteinase（ MMP） 2 and cyclin D1 at mRNA and protein levels was determined by real-time PCR and Western blot.After induction of the cells by recombinant Shh proteins,the changes of angiogenesis and proliferation ability in each group were detected.At the same time,the transformation of related gene was examined.RESULTS：Compared with control group,the angiogenic ability in interfered group was decreased,and no obvious difference of proliferation ability was observed.The expression of VEGF-A and MMP2 was declined significantly,and the expression of cyclin D1 was not obviously changed.Recombinant Shh proteins improved the expression of FOXQ1 gene.Compared with NC-shRNA group,after induction,the angiogenic ability of FOXQ1-shRNA group was decreased,and the proliferation ability was not obviously changed.CONCLUSION：FOXQ1 gene mediates the angiogenic ability but does not affect the proliferation ability of SW480 cells.Meanwhile,it may be regulated by shh pathway.