The orphan nuclear receptor NUR77 promotes trophoblast invasion at early pregnancy through paracrine placental growth factor
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NR4A1 (NUR77) is an orphan nuclear receptor that has been implicated in both cell survival and apoptosis. However, the role of NUR77 in trophoblast function during early placenta development has not been fully elucidated. In this study, we showed that NUR77 expression was significantly lower in the villi of the recurrent miscarriage (RM) group compared to that in the healthy controls (HCs) group. We used immunohistochemistry and found that NUR77 was highly expressed in human placental villi during early pregnancy, especially in syncytiotrophoblast (STB), and was expressed at a much lower level in STB from the RM group than in those from HC group. Western blotting data further confirmed that NUR77 was highly expressed in primary human term placental STB and the FSK-induced BeWo cell line. Moreover, antibody array screening and ELISA revealed that NUR77 promoted significant placental growth factor (PGF) expression during trophoblast fusion. Ectopic overexpression and knockdown experiments demonstrated that PGF was a novel downstream target of NUR77, and serum PGF expression correlated positively with trophoblast NUR77 mRNA levels in HCs and RM patients. Importantly, bioinformatics analysis identified two NUR77 binding sites in the PGF promoter region, and chromatin immunoprecipitation (ChIP) coupled with Western blotting analysis further verified that NUR77 bound directly to the PGF promoter region and promoted PGF expression. Furthermore, in a BeWo/HTR-8 co-culture system, FSK-induced BeWo-secreted PGF promoted HTR-8 cell migration and invasion, and an anti-PGF antibody reversed this effect. Collectively, these results indicated that NUR77 may play a key role in regulating trophoblast invasion at early pregnancy.
NUR77 expression was significantly decreased in the syncytiotrophoblast of the recurrent miscarriage group compared to that in the healthy control group.
NUR77 promoted PGF expression during trophoblast fusion.
ChIP and western blotting experiments verified that NUR77 bound directly to the PGF promoter region and activated PGF expression in trophoblast.
Trophoblast-derived PGF promoted HTR-8 cell migration and invasion in a cell co-culture system.
KeywordsNUR77 PGF Trophoblast invasion Syncytiotrophoblast Early pregnancy
X.-C.L., X.-J.Y., and F. -J.T. performed the experiments; W. H, F. J, and J.L. contributed to the statistical analyses; P.-Y.W. provided material support; Y.-M. W. contributed valuable advice to the development of the manuscript; X.-C.L., X.-J.Y., and F.-J.T. wrote the manuscript; and F.-J.T. designed the study and directed the project.
The work was supported by the National Natural Science Foundation of China (81401218 to F.-J.T.) and by the Family Planning Project of the Shanghai Health and Family Planning Commission (20164037 to X.-C.L.), the key projects of the Shanghai Municipal Health and Family Planning Commission (201640012 to F.-J.T.), and the Shanghai Jiao Tong University Medicine-Engineering Fund (YG2017 MS38 to F.-J.T.).
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Conflict of interest
The authors declare that they have no conflict of interest.
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