Effects of Notch2 and Notch3 on Cell Proliferation and Apoptosis of Trophoblast Cell Lines

Abstract

Aims: To investigate the effect of Notch2 and Notch3 on cell proliferation and apoptosis of two trophoblast cell lines, BeWo and JAR.Methods: Notch2 and Notch3 expression in BeWo and JAR cells was upregulated or downregulated using lentivirus-mediated overexpression or RNA interference. The effect of Notch2 and Notch3 on cell proliferation was assessed by the CCK-8 assay. The effect of Notch2 and Notch3 on the apoptosis of BeWo and JAR cells was evaluated by flow cytometry using the Annexin V-PE Apoptosis kit. Lentivirus-based overexpression vectors were constructed by cloning the full-length coding sequences of human Notch2 and Notch3 C-terminally tagged with GFP or GFP alone (control) into a lentivirus-based expression vector. Lentivirus-based gene silencing vectors were prepared by cloning small interfering sequences targeting human Notch2 and Notch3 and scrambled control RNA sequence into a lentivirus-based gene knockdown vector. The effect of Notch2 and Notch3 on cell proliferation was assessed by the CCK-8 assay. And the effect of Notch2 and Notch3 on the apoptosis of BeWo and JAR cells was evaluated by flow cytometry using the Annexin V PE Apoptosis kit.Results: We found that the downregulation of Notch2 and Notch3 gene expression in BeWo and JAR cells resulted in an increase in cell proliferation, while upregulation of Notch3 and Notch2 expression led to a decrease in cell proliferation. Moreover, the overexpression of Notch3 and Notch2 in BeWo and JAR cells reduced apoptosis in these trophoblast cell lines, whereas apoptosis was increased in the cells in which the expression of Notch3 and Notch2 was downregulated.Conclusions: Notch2 and Notch3 inhibited both cell proliferation and cell apoptosis in BeWo and JAR trophoblast cell lines.

Notch2 and Notch3 suppress the proliferation and mediate invasion of trophoblast cell lines

“…Previous research showed that Notch2 expression decreased, and Notch3 was overexpressed, in the placentas of patients with preeclampsia compared with normal placentas ( Zhao et al, 2014 ), which indicate that Notch2 and Notch3 may play important roles in the pathology of preeclampsia. In our previously published research, we demonstrated that Notch2 and Notch3 exert antiproliferative effects on trophoblast cells ( Zhao et al, 2015 ). In the present study, we present more evidence that high levels of Notch2 and Notch3 are able to induce cell cycle arrest at G2/M and G0/G1 phases, subsequently resulting in suppressed proliferation.…”

“…Our previously reported results indicated that the expression of Notch2 and Notch3 is associated with the proliferation of BeWo and JAR cells ( Zhao et al, 2015 ). Specifically, Notch2 knockdown significantly promoted BeWo cell proliferation, while Notch3 overexpression decreased the growth rate of BeWo cells, compared with that of the negative control.…”

“…The results of a previous study showed an obvious upregulation of Notch3 expression, and a downregulation of Notch2 expression, in placentas from patients with early onset severe preeclampsia compared with those in normal placentas ( Zhao et al, 2014 ). Notch2 and Notch3 inhibit both cell proliferation and cell apoptosis in BeWo and JAR cells ( Zhao et al, 2015 ). However, the mechanisms by which Notch2 and Notch3 are related to preeclampsia remain unknown.…”

MicroRNA‐16 is involved in the pathogenesis of pre‐eclampsia via regulation of Notch2

“…Furthermore, Zhao et al (2015) have suggested that Notch2 and Notch3 repressed proliferation and apoptosis in cell lines of BeWo and JAR trophoblasts. It was conveyed by a previous study that Notch signaling is a critical part of the process of invasion of fetal trophoblast cell and reconstruction of maternal blood vessels (Hunkapiller et al, 2011).…”

HIF1α‐mediated AIMP3 suppression delays stem cell aging via the induction of autophagy

“…AIMP3 was induced in both si‐HIF1α‐ and si‐Hey1‐treated hpMSCs, and the synergistic accumulation of AIMP3 was observed in both si‐HIF1α‐ and si‐Hey1‐treated cells, indicating that both HIF1α and Hey1 negatively regulate AIMP3 in an additive manner (Figure b). Because Notch3, one of the Notch receptors, suppresses the proliferation of trophoblasts in the placenta and functions as a tumor suppressor by inducing cellular senescence (Cui, Kong, Xu, & Zhang, ; Zhao, Zhuang, Huang, Feng, & Lin, ), the differential expression of Notch3 was first examined under different oxygen levels. Under hypoxic conditions, the levels of Notch3 at the middle and late passages were repressed compared to those under normoxia (Figure c), and the forced suppression of Notch3 led to the significant repression of AIMP3 and decreased p16 INK4a (Figure d), indicating that Notch3 is able to induce senescence via AIMP3 upregulation.…”