全反式维甲酸通过调控sFlt-1表达对滋养层细胞侵袭及促血管形成能力的影响

Effects of ATRA on trophoblast cell invasion and angiogenesis by regulating sFlt-1 expression

目的探讨全反式维甲酸(ATRA)对可溶性血管内皮生长因子受体1(sFlt-1)的调控作用及其对人胎盘滋养层细胞侵袭及促血管形成能力的影响。方法采用0.15μmol/L ATRA处理HTR-8/SVneo细胞12、24、48、72 h,RT-PCR和ELISA分别检测细胞中sFlt-1 mRNA以及上清液中sFlt-1蛋白水平。采用携带sFlt-1过表达载体慢病毒感染HTR-8/SVneo细胞,并采用0.15μmol/L ATRA处理24 h,采用ELISA检测细胞上清液中sFlt-1、胎盘生长因子(PIGF)和血管内皮生长因子(VEGF)的蛋白含量;Transwell实验检测细胞的侵袭和迁移能力;小管形成实验检测细胞体外促血管形成能力;Western blot检测细胞中基质金属蛋白酶-2(MMP-2)和MMP-9蛋白表达水平。结果 ATRA可呈时间依赖性的抑制HTR-8/SVneo细胞中sFlt-1 mRNA表达及其蛋白的分泌;ATRA干预后,HTR-8/SVneo细胞上清液中sFlt-1蛋白水平降低(P <0.05),PIGF和VEGF蛋白水平升高(P <0.05),而细胞侵袭和迁移能力、血管成管数目以及MMP-9和MMP-2蛋白表达水平均增加(P <0.05)。然而,sFlt-1过表达可抑制ATRA对HTR-8/SVneo细胞的干预效果。结论ATRA可通过抑制sFlt-1表达增强滋养层细胞侵袭及促血管形成能力。

Objective To investigate the regulatory effect of all-trans retinoic acid(ATRA) on soluble vascular endothelial growth factor receptor 1(sFlt-1) and its effect on the invasion and angiogenesis of human placental trophoblast cells.Methods HTR-8/SVneo cells were treated with 0.15 μM ATRA for 12,24,48 and 72 h,then the expression levels of sFlt-1 mRNA and protein were detected by RT-PCR and ELISA.HTR-8/SVneo cells were infected with sFlt-1 overexpresses lentivirus,and treated with 0.15 μmol/L ATRA for 24 h.The protein contents of sFlt-1,PIGF and VEGF in the cell culture supernatant were detected by ELISA.The abilities of invasion and migration were detected by transwell assay.The pro-angiogenesis ability of cells in vitro was tested by tubule formation assay.The protein expression levels of MMP-2 and MMP-9 were detected by Western blot.Results ATRA inhibited the mRNA expression and protein secretion of sflt-1 in HTR-8/SVneo cells in a time-dependent manner;After ATRA treatment,the content of sFlt-1 protein in the cell supernatant of HTR-8/SVneo cells was decreased(P <0.05),and the protein contents of PIGF and VEGF were increased(P <0.05),the abilities of invasion and migration,the number of vascularization tubes and the protein expression levels of MMP-9 and MMP-2 were increased(P <0.05).However,overexpression of sFLT-1 inhibited the intervention effect of ATRA on HTR-8/SVneo cells.Conclusion ATRA enhances trophoblast cell invasion and angiogenesis by inhibiting sflt-1 expression.