The Affiliated Wuxi Maternity and Child Health Care Hospital of Nanjing Medical University, Huaishuxiang Road, Chong an Street, Wuxi, 214002, China.
Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, 536 Changle Road, Shanghai, 200040, China.
Placenta. 2019 May;80:27-35. doi: 10.1016/j.placenta.2019.03.001. Epub 2019 Mar 11.
This study aimed to test the influence of homeobox B7 (HoxB7) on the proliferation, invasion, and migration of human trophoblast cells and to reveal the down-regulation of HoxB7 on the transcriptional suppression of Dick Kopf-related protein1 (DKK1) and of Cysteine-rich glycosylated wingless protein 1 (Wnt1)/β-catenin in intrauterine fetal growth retardation (FGR).
Quantitative measurement of HoxB7, DKK1, Wnt1, and β-catenin was performed in human placentas collected from normal pregnancies and from FGR with quantitative real time PCR (qRT-PCR). Cultured HTR-8/SVneo cells, transfected with a lentiviral plasmid that in-frame expresses human HoxB7 gene, were applied to functional assessment to study the biological impact of HoxB7 gene on DKK1, Wnt1, and β-catenin. Counting Kit-8, Transwell invasion assays, and flow cytometry were applied for the functional measurements.
The expression of HoxB7 was significantly increased, and of DKK1, Wnt1, and β-catenin was decreased, in FGR placenta tissues and in HTR-8/SVneo cells. Function studies revealed that overexpression of HoxB7 inhibited proliferation, migration, and invasion in HTR-8/SVneo cells. DKK1, Wnt1, and β-catenin were down-regulated in HTR-8/SVneo cells, inversely correlated with HoxB7 expression. Overexpression of HoxB7 showed a suppressive effect on proliferation, migration, and invasion in the HTR-8/SVneo cells.
Our results indicate that HoxB7 inhibited human trophoblast cell differentiation by down-regulating DKK1 expression and that it may affect transcription of Wnt1/β-catenin. The activation of HoxB7 might suppress the cell differentiation in HTR-8/SVneo cell cultures. The Wnt/β-catenin signaling pathway may play a significant role in the pathogenesis of FGR by regulating the invasion and proliferation of trophoblasts.
本研究旨在检测同源盒 B7(HoxB7)对人滋养层细胞增殖、侵袭和迁移的影响,并揭示 HoxB7 下调对 Dickkopf 相关蛋白 1(DKK1)和富含半胱氨酸糖基化无翅型蛋白 1(Wnt1)/β-连环蛋白转录抑制的作用,这些作用与宫内胎儿生长迟缓(FGR)有关。
采用实时定量 PCR(qRT-PCR)法检测正常妊娠和 FGR 胎盘组织中 HoxB7、DKK1、Wnt1 和 β-连环蛋白的含量。应用慢病毒质粒转染滋养层细胞株 HTR-8/SVneo,使其过表达人 HoxB7 基因,进行功能评估,以研究 HoxB7 基因对 DKK1、Wnt1 和 β-连环蛋白的生物学影响。采用细胞计数试剂盒-8(CCK-8)法、Transwell 侵袭实验和流式细胞术进行功能检测。
FGR 胎盘组织和 HTR-8/SVneo 细胞中 HoxB7 表达明显增加,而 DKK1、Wnt1 和 β-连环蛋白表达降低。功能研究表明,HoxB7 过表达抑制 HTR-8/SVneo 细胞增殖、迁移和侵袭。HTR-8/SVneo 细胞中 DKK1、Wnt1 和 β-连环蛋白下调,与 HoxB7 表达呈负相关。HoxB7 过表达对 HTR-8/SVneo 细胞的增殖、迁移和侵袭具有抑制作用。
本研究结果表明,HoxB7 通过下调 DKK1 的表达抑制人滋养层细胞的分化,可能影响 Wnt1/β-连环蛋白的转录。HoxB7 的激活可能抑制 HTR-8/SVneo 细胞培养物中的细胞分化。Wnt/β-连环蛋白信号通路可能通过调节滋养细胞的侵袭和增殖在 FGR 的发病机制中发挥重要作用。