SFRP2 suppresses trophoblast cell migration by inhibiting the Wnt/β‑catenin pathway

Abstract

The present study investigates the role of Secreted Frizzled‑Related Protein 2 (SFRP2) in trophoblast cells, a key factor in preeclampsia (PE) progression. Elevated levels of Secreted Frizzled‑Related Protein 1/3/4/5 (SFRP1/3/4/5) are associated with PE, but the role of SFRP2 is unclear. We analyzed SFRP2 expression in PE placental tissue using the GSE10588 dataset and overexpressed SFRP2 in JEG‑3 cells via lentiviral transfection. The viability, migration, apoptosis, and proliferation of SFRP2‑overexpressing JEG‑3 cells were assessed using Cell Counting Kit‑8, Transwell assays, flow cytometry, and EdU staining. Additionally, we evaluated the impact of SFRP2 overexpression on key proteins in the Wnt/β‑catenin pathway and apoptosis markers (Bax, cleaved‑caspase 3, BCL‑2, MMP9, E‑cadherin, Wnt3a, Axin2, CyclinD1, c‑Myc, p‑β‑catenin, β‑catenin, phosphorylated Glycogen Synthase Kinase 3 beta (p‑GSK3β), and GSK3β) through western blotting. Results showed high SFRP2 mRNA and protein expression in PE placenta and JEG‑3 cells post‑transfection. SFRP2 overexpression significantly reduced JEG‑3 cell viability, proliferation, and migration, while increasing apoptosis. It also altered expression levels of Wnt pathway proteins, suggesting SFRP2's potential as a therapeutic target for PE by inhibiting trophoblast cell migration through the Wnt/β‑catenin signaling cascade.

Keywords: Wnt; migration; preeclampsia; secreted frizzled‑related protein 2.

Figure 1.

SFRP2 is upregulated in JEG-3 cells. (A) GSE10588 dataset demonstrated upregulated expression of SFRP2 in PE placental tissue compared with that in Nor pregnancy. mRNA and protein expression of SFRP2 in JEG-3 cells was determined by (B) quantitative PCR and (C) western blotting. Grayscale analysis was performed by ImageJ software. *P<0.05, ***P<0.001 vs. NC. PE, preeclampsia; Nor, normal; NC, negative control; OE, overexpression; SFRP2, secreted frizzled-related protein 2.

Figure 2.

Elevated SFRP2 inhibits viability and migration while increasing apoptosis of trophoblast cells. (A) Viability of JEG-3 cells was detected by Cell Counting Kit-8. (B) Migration of JEG-3 cells was determined by Transwell assay. Magnification, ×100. (C) Ratio of apoptosis of JEG-3 cells on flow cytometric detection. ***P<0.001. NC, negative control; OE, overexpression; SFRP2, secreted frizzled-related protein 2; OD, optical density.

Figure 3.

SFRP2 overexpression inhibits migration and promotes apoptosis of trophoblast cells. (A) JEG-3 cell proliferation was determined using EdU staining assay. (B) Protein expression of Bax, cleav-caspase 3, BCL-2, MMP9 and E-cadherin by western blotting. Grayscale analysis was performed by ImageJ software. ***P<0.001. NC, negative control; OE, overexpression; SFRP2, secreted frizzled-related protein 2; cleav, cleaved; M, marker.

Figure 4.

High SFRP2 expression attenuates Wnt signaling. (A) Protein expression of Wnt3a, Axin2, CyclinD1, c-Myc and β-catenin verified by western blot. (B) Grayscale analysis was performed by ImageJ software. (C) Protein levels of p-β-catenin, p-GSK3β and GSK3β in OE-SFRP2 JEG-3 cells verified by western blot. **P<0.05, ***P<0.001. NC, negative control; OE, overexpression; SFRP2, secreted frizzled-related protein 2; p-, phosphorylated; GSK, Glycogen Synthase Kinase 3β; M, protein Marker.