miR-663调控MMP11对妊娠滋养层细胞侵袭和迁移的影响

Effects of miR-663 regulates MMP11 on the invasion and migration of gestational trophoblast cells

目的探讨微小RNA-663(miR-663)调控基质金属蛋白酶11(MMP11)对妊娠滋养层细胞侵袭和迁移的影响。方法收集2018年7月至2020年7月重庆医科大学附属第一医院綦江医院收治的妊娠期子痫前期患者行剖宫产留取的胎盘组织19例,同时收集健康妊娠女性行剖宫产留取的胎盘组织19例(对照组织)。采用qRT-PCR检测子痫前期患者胎盘组织及对照组织中miR-663和MMP11 mRNA表达。在妊娠滋养层JEG-3细胞中转染MMP11过表达载体、miR-663抑制剂或/和MMP11小干扰RNA,采用CCK-8法测定增殖,Transwell小室测定迁移和侵袭。生物学信息学软件预测miR-663的靶基因,荧光素酶报告系统鉴定二者的靶向关系。结果与对照组织比较,子痫前期患者胎盘组织中miR-663表达水平升高(P<0.05),MMP11 mRNA表达水平降低(P<0.05)。下调miR-663或过表达MMP11均能够提高妊娠滋养层细胞的增殖活性,促进细胞迁移和侵袭。抑制MMP11能够逆转下调miR-663对妊娠滋养层细胞增殖、侵袭和迁移的促进作用。miR-663靶向负调控MMP11。结论下调miR-663调控MMP11会促进妊娠滋养层细胞的侵袭和迁移。

Objective To investigate the effect of microRNA-663(miR-663)regulating matrix metalloproteinase11(MMP11)on the invasion and migration of gestational trophoblast cells.Methods A total of 19 cases of placental tissue taken by cesarean section from pregnant preeclampsia patients admitted to Qijiang Hospital,the First Affiliated Hospital of Chongqing Medical University from July 2018 to July 2020 were collected,as well as 19 cases of placental tissue taken by cesarean section from healthy pregnant women(control tissue).qRT-PCR method was used to detect the expression of miR-663 and MMP11 mRNA in the placenta of patients with preeclampsia and control tissue.MMP11 overexpression vector,miR-663 inhibitor or/and MMP11 small interfering RNA were transfected into gestational trophoblast JEG-3 cells,and CCK-8 method was used to measure proliferation,Transwell chamber was used to measure migration and invasion.The bioinformatics software was used to predict the target genes of miR-663,and the luciferase reporter system was used to identify the target relationship between the two.Results Compared with control tissues,the expression level of miR-663 in the placenta tissue of patients with preeclampsia increased(P<0.05),and the expression levels of MMP11 mRNA decreased(P<0.05).Down-regulation of miR-663 or over-expression of MMP11 can increase the proliferation activity of gestational trophoblast cells and promote cell migration and invasion.Inhibiting MMP11 can reverse the promotion of down-regulating miR-663 on the proliferation,invasion and migration of gestational trophoblast cells.miR-663 targeted the negative regulation of MMP11.Conclusion Down-regulating miR-663 regulates MMP11 can promote the invasion and migration of gestational trophoblast cells.