过表达NDRG1通过调控VEGF/sFlt-1轴增强缺氧条件下滋养细胞的促血管形成能力  

Overexpression of NDRG1 enhances the pro-angiogenesis ability of trophoblast cells under hypoxic by regulating VEGF/sFlt-1 axis

作　　者：何宜静 欧琼 李婷 刘珏 He Yijing;Ou Qiong;Li Ting(Dept of Obstetrics and Gynecology,The Second Affiliated Hospital of Nanhua University,Hengyang 421001)

机构地区：[1]南华大学附属第二医院妇产科,衡阳421001

出　　处：《安徽医科大学学报》2020年第5期745-750,共6页Acta Universitatis Medicinalis Anhui

基　　金：湖南省卫生健康委2019年度科研计划项目(编号:B2019116)。

摘　　要：目的探讨缺氧条件下N-myc下游调节基因1(NDRG1)过表达对滋养细胞HTR-8/SVneo促血管形成能力的影响。方法构建NDRG1过表达慢病毒载体并感染HTR-8/SVneo细胞,建立NDRG1基因稳定过表达的HTR-8/SVneo细胞株,随后采用低氧(1%O2)干预24 h。qRT-PCR检测缺氧后细胞中NDRG1、血管内皮生长因子(VEGF)和可溶性血管内皮生长因子受体1(sFlt-1)mRNA表达水平;Western blot检测细胞中NDRG1、基质金属蛋白酶-2(MMP-2)和MMP-9等蛋白表达水平;ELISA法检测细胞培养上清液中VEGF和sFlt-1蛋白含量;小管形成实验检测细胞体外促血管形成能力;Transwell实验检测细胞侵袭能力。结果缺氧可诱导HTR-8/SVneo细胞中NDRG1、VEGF、sFlt-1等mRNA表达,并促进NDRG1蛋白表达及VEGF和sFlt-1蛋白的分泌,还可抑制MMP-2、MMP-9等蛋白表达以及降低细胞体外促血管形成能力和侵袭能力。NDRG1过表达可促进低氧条件下HTR-8/SVneo细胞VEGF mRNA的表达和蛋白的分泌,同时抑制sFlt-1 mRNA表达及其蛋白的分泌。此外,NDRG1过表达还可上调低氧条件下HTR-8/SVneo细胞中MMP-2和MMP-9蛋白表达水平,并促进细胞体外血管形成能力及侵袭能力。结论NDRG1过表达可增强缺氧条件下滋养细胞的促血管形成能力和侵袭能力,其机制可能与促进VEGF表达和抑制sFlt-1表达有关。Objective To investigate the effect of N-myc downstream regulatory gene 1(NDRG1)overexpression on the pro-angiogenic ability of trophoblastic cells HTR-8/SVneo under hypoxic condition.Methods HTR-8/SVneo cells with stable overexpression of NDRG1 were established by infecting with NDRG1 overexpression lentivirus,and then hypoxia(1%O2)was used to intervene for 24 h.qRT-PCR was used to detect the mRNA expression levels of NDRG1,vascular endothelial growth factor(VEGF)and solunle vascular endothelial growth factor receptor-1(sFlt-1).The protein expression levels of NDRG1,MMP-2 and MMP-9 were detected by Western blot.The protein contents of VEGF and sFlt-1 in the cell culture supernatant were detected by ELISA.The ability of promoting pro-angiogenesis in vitro was tested by tubule formation assay.The invasion ability of cells was detected by transwell assay.Results Hypoxia could induce the mRNAs expression of NDRG1,VEGF and sFlt-1 in HTR-8/SVneo cells,promote the protein expression of NDRG1 and the protein secretions of VEGF and sFlt-1,inhibit the proteins expression of MMP-2 and MMP-9,and reduce the pro-angiogenesis and invasion ability of cells in vitro.Overexpression of NDRG1 promoted the mRNA expression and protein secretion of VEGF in HTR-8/SVneo cells under hypoxic condition,and inhibited the mRNA expression and protein secretion of sFlt-1.In addition,overexpression of NDRG1 also increased the protein expression levels of MMP-2 and MMP-9 in HTR-8/SVneo cells under hypoxic condition,and promoted the pro-angiogenesis and invasion ability of cells in vitro.Conclusion Overexpression of NDRG1 can enhance the pro-angiogenesis and invasion ability of trophoblast cells under hypoxic condition,and its mechanism may be related to the promotion of VEGF expression and inhibition of sFlt-1 expression.

关 键 词：N-myc下游调节基因1 滋养细胞 低氧 血管形成 侵袭 

分 类 号：R714.244[医药卫生—妇产科学]