HOXA9 transcriptionally regulates the EPHB4 receptor to modulate trophoblast migration and invasion.

Functional placenta formation is crucially dependent on extravillous trophoblast migration and invasion. EPHB4 has been identified to play a negative but important role in regulating trophoblast biological function, whereas the upstream regulation mechanism remains unknown. As reported, there is a transcriptional stimulation of EPHB4 expression consequent to HOXA9 activation in endothelial cells (ECs). Therefore, this study is conducted to investigate the role of HOXA9 and its relationship with EPHB4 in trophoblast cells. Both mRNA and protein expression levels of HOXA9 and EPHB4 were measured in preeclamptic placenta (n = 15) and normal placenta (n = 15). Next, the expression and location of HOXA9 and EPHB4 in first-trimester villi were shown via immunohistochemistry. Trophoblast cell line HTR-8/SVneo was used to explore the effect of HOXA9 on EPHB4 expression and trophoblast bioactivity by gain- and loss-of function studies. In addition, chromatin immunoprecipitation (ChIP) and luciferase assays were conducted to clarify the regulation mechanism of HOXA9 on EPHB4 expression in HTR-8/SVneo. HOXA9 and EPHB4 expression were increased in preeclamptic placenta compared with normal placenta. HOXA9 could promote EPHB4 expression and impaired HTR-8/SVneo cells migration and invasion. ChIP and luciferase assays revealed that HOXA9 could directly bind to EPHB4 promoter and promoted its transcription. HOXA9 transcriptionally regulated EPHB4 expression to modulate trophoblasts migration and invasion, which may suggest a contribution of HOXA9-EPHB4 in the poor placentation in the pathogenesis of preeclampsia.