Role of peroxiredoxin2 downregulation in recurrent miscarriage through regulation of trophoblast proliferation and apoptosis

Abstract

Peroxiredoxin (Prdx) 2 is an antioxidant protein that utilizes its redox-sensitive cysteine groups to reduce hydrogen peroxide molecules and protect cells against oxidative damage from reactive oxygen species (ROS). However, its function in trophoblasts at the maternal–fetal interface has not been clarified yet. In this study, significantly lower Prdx2 expression was found in the first-trimester villous cytotrophoblasts of patients with recurrent miscarriage (RM) than in cytotrophoblasts from healthy controls. Further, Prdx2 knockdown inhibited proliferation and increased apoptosis of trophoblast cells. The reason for this may be an increase in the level of cellular ROS after knockdown of Prdx2, which may subsequently lead to an increase in the expression of phosphorylated p53 (p-p53) and p38-MAPK/p21. Prdx2 knockdown also impaired the fusion of BeWo cells induced by forskolin. Bioinformatics analysis identified a c-Myc-binding site in the Prdx2 promoter region, and chromatin immunoprecipitation verified that c-Myc directly bound to a site in this locus. Suppression and overexpression of c-Myc resulted in reduction and increase of Prdx2 expression respectively. Furthermore, we demonstrated that c-Myc was downregulated in the first-trimester cytotrophoblasts of patients with RM, and its downregulation is also related with inhibited cell proliferation, increased apoptosis, as well as upregulated p21 expression and p-p53/p53 ratio. Our findings indicate that Prdx2 might have an important role in the regulation of trophoblast proliferation and apoptosis during early pregnancy, and that its expression is mediated by c-Myc. Thus, these two proteins may be involved in the pathogenesis of RM and may represent potential therapeutic targets.

SPRY4 regulates trophoblast proliferation and apoptosis via regulating IFN‐γ‐induced STAT1 expression and activation in recurrent miscarriage

“…Data are expressed as the mean ± SEM*P < .05, **P < .01, ***P < .001, ****P < .0001, ns: not significant (ROS) are necessary for cell growth signaling during normal pregnancy, high levels of hypoxia-induced ROS lead to a transient or long-term cell cycle arrest and are correlated with various placental pathologies including RM, PE, and IUGR. 4 In accordance with its elevation in response to hypoxia, we observed that the expression of SPRY4 was increased in the first-trimester trophoblasts from patients with RM. Furthermore, in vitro experiments showed that SPRY4 overexpression restrained cell proliferation, impaired DNA synthesis, and accelerated apoptosis in the HTR8 cell line, while SPRY4 deficiency reversed these effects.…”

“…40 Trophoblast dysfunctions resulted from the unbalanced level of trophoblast proliferation and apoptosis are critical factors to the development of RM. 4 In the past decade, SPRY genes have been considered as a general inhibitor of RTK-dependent signaling pathways, acting as tumour suppressors in several cancers via regulating cell fate.…”

“…Immunohistochemical (IHC) staining of human villous tissue was performed as previously described. 4 Briefly, the paraffin-embedded tissue slides were dewaxed and rehydrated and antigens were retrieved in boiling EDTA buffer. After incubation with 3% hydrogen peroxide and blockage with 5% BSA, the sections were incubated with primary Abs: anti-SPRY4 (1:50 dilution, Proteintech), anti-…”

“…2 Defective abilities of CTBs have been proven to result in shallow placental implantation, and many pregnancy-related complications such as preeclampsia (PE), intrauterine growth retardation (IUGR) and recurrent miscarriage (RM) are partially construed as a consequence of impaired trophoblast biological behaviors. 3,4 RM currently refers to two or more consecutive spontaneous losses until 20 weeks of gestation, frustrating 1-5% of couples of childbearing age. 5 Basically, genetic defects, anatomical abnormalities, endocrine disturbances, infections, immune dysfunctions, thrombotic disorders, and unexplained causes are implicated in the occurrence of RM.…”

Derivation of macaque trophoblast stem cells

“…Unlike human embryonic ESCs, macaque ESCs do not respond to BMP4 treatment to promote differentiation to trophectoderm 58,59 , however, it is plausible that this cluster represents established TSCs that are positioned for differentiation. A more conservative analysis, K-means analysis, revealed a distinct axis of up or down-regulated genes between clusters 1 and 2, including NUCB2, APOE, PRDX2, and LDHA, that have previously been shown to have roles in trophoblast function [60][61][62][63][64] .…”