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MSX2 Induces Trophoblast Invasion in Human Placenta.

Liang H, Zhang Q, Lu J, Yang G, Tian N, Wang X, Tan Y, Tan D - PLoS ONE (2016)

Bottom Line: Furthermore, we found that the in vitro invasive ability of HTR8/SVneo cells was enhanced by exogenous overexpression of MSX2, and that this effect was accompanied by increased protein expression of matrix metalloproteinase-2 (MMP-2), vimentin, and β-catenin.Meanwhile, overexpression of MSX2 and treatment with the MSX2-specific siRNA resulted in decreased and increased E-cadherin expression, respectively, in JEG-3 cells.Collectively, our results suggest that MSX2 may induce human trophoblast cell invasion, and dysregulation of MSX2 expression may be associated with pre-eclampsia.

View Article: PubMed Central - PubMed

Affiliation: Laboratory Animal Center, Chongqing Medical University, Chongqing, China.

ABSTRACT
Normal implantation depends on appropriate trophoblast growth and invasion. Inadequate trophoblast invasion results in pregnancy-related disorders, such as early miscarriage and pre-eclampsia, which are dangerous to both the mother and fetus. Msh Homeobox 2 (MSX2), a member of the MSX family of homeobox proteins, plays a significant role in the proliferation and differentiation of various cells and tissues, including ectodermal organs, teeth, and chondrocytes. Recently, MSX2 was found to play important roles in the invasion of cancer cells into adjacent tissues via the epithelial-mesenchymal transition (EMT). However, the role of MSX2 in trophoblastic invasion during placental development has yet to be explored. In the present study, we detected MSX2 expression in cytotrophoblast, syncytiotrophoblast, and extravillous cytotrophoblast cells of first or third trimester human placentas via immunohistochemistry analysis. Furthermore, we found that the in vitro invasive ability of HTR8/SVneo cells was enhanced by exogenous overexpression of MSX2, and that this effect was accompanied by increased protein expression of matrix metalloproteinase-2 (MMP-2), vimentin, and β-catenin. Conversely, treatment of HTR8/SVneo cells with MSX2-specific siRNAs resulted in decreased protein expression of MMP-2, vimentin, and β-catenin, and reduced invasion levels in a Matrigel invasion test. Notably, however, treatment with the MSX2 overexpression plasmid and the MSX2 siRNAs had no effect on the mRNA expression levels of β-catenin. Meanwhile, overexpression of MSX2 and treatment with the MSX2-specific siRNA resulted in decreased and increased E-cadherin expression, respectively, in JEG-3 cells. Lastly, the protein expression levels of MSX2 were significantly lower in human pre-eclamptic placental villi than in the matched control placentas. Collectively, our results suggest that MSX2 may induce human trophoblast cell invasion, and dysregulation of MSX2 expression may be associated with pre-eclampsia.

No MeSH data available.


Related in: MedlinePlus

MSX2 is significantly downregulated in the placental villi from PE patients.(A) Immunostaining of MSX2 in the EVTs from normal and PE placentas at similar gestational age (a and b). CK7 was used as a marker for EVTs (c and d). Negative control images (NEG) in which normal IgG was used to replace the primary antibody showed no specific positive staining (e and f). Boxed region is enlarged on the under panel. Scale bar = 200μm. NP: nomal term placenta PE: pre-eclampsia (B) Western blot analysis of MSX2 expression in the human placental villi from term placentas (38 weeks; n = 3) and PE placentas (38 weeks; n = 3). (C) Results obtained in B were quantified by measuring the protein band intensity of MSX2 relative to that of GAPDH (n = 8; t-test; **P < 0.01).
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pone.0153656.g007: MSX2 is significantly downregulated in the placental villi from PE patients.(A) Immunostaining of MSX2 in the EVTs from normal and PE placentas at similar gestational age (a and b). CK7 was used as a marker for EVTs (c and d). Negative control images (NEG) in which normal IgG was used to replace the primary antibody showed no specific positive staining (e and f). Boxed region is enlarged on the under panel. Scale bar = 200μm. NP: nomal term placenta PE: pre-eclampsia (B) Western blot analysis of MSX2 expression in the human placental villi from term placentas (38 weeks; n = 3) and PE placentas (38 weeks; n = 3). (C) Results obtained in B were quantified by measuring the protein band intensity of MSX2 relative to that of GAPDH (n = 8; t-test; **P < 0.01).

Mentions: Inadequate trophoblast migration/invasion and impaired spiral artery remodeling can result in poor placental perfusion leading to pregnancy-related diseases such as PE [23]. Placental villi from pre-eclamptic and normal pregnancy patients of close gestational stages were analyzed by immunohistochemistry (Fig 7A) and were dissected and subjected to western blot analysis (Fig 7B). Immunohistochemistry showed weak positive signals for MSX2 in the EVTs from PE placentas (Fig 7Ab) compared with the strong MSX2 staining in the EVTs from normal placentas (Fig 7Aa). Western blotting(Fig 7B) and statistical analyses o(Fig 7C) indicated that MSX2 was expressed at significantly lower levels in the placental villi from pre-eclamptic patients than in those from control placentas (n = 8).


MSX2 Induces Trophoblast Invasion in Human Placenta.

Liang H, Zhang Q, Lu J, Yang G, Tian N, Wang X, Tan Y, Tan D - PLoS ONE (2016)

MSX2 is significantly downregulated in the placental villi from PE patients.(A) Immunostaining of MSX2 in the EVTs from normal and PE placentas at similar gestational age (a and b). CK7 was used as a marker for EVTs (c and d). Negative control images (NEG) in which normal IgG was used to replace the primary antibody showed no specific positive staining (e and f). Boxed region is enlarged on the under panel. Scale bar = 200μm. NP: nomal term placenta PE: pre-eclampsia (B) Western blot analysis of MSX2 expression in the human placental villi from term placentas (38 weeks; n = 3) and PE placentas (38 weeks; n = 3). (C) Results obtained in B were quantified by measuring the protein band intensity of MSX2 relative to that of GAPDH (n = 8; t-test; **P < 0.01).
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pone.0153656.g007: MSX2 is significantly downregulated in the placental villi from PE patients.(A) Immunostaining of MSX2 in the EVTs from normal and PE placentas at similar gestational age (a and b). CK7 was used as a marker for EVTs (c and d). Negative control images (NEG) in which normal IgG was used to replace the primary antibody showed no specific positive staining (e and f). Boxed region is enlarged on the under panel. Scale bar = 200μm. NP: nomal term placenta PE: pre-eclampsia (B) Western blot analysis of MSX2 expression in the human placental villi from term placentas (38 weeks; n = 3) and PE placentas (38 weeks; n = 3). (C) Results obtained in B were quantified by measuring the protein band intensity of MSX2 relative to that of GAPDH (n = 8; t-test; **P < 0.01).
Mentions: Inadequate trophoblast migration/invasion and impaired spiral artery remodeling can result in poor placental perfusion leading to pregnancy-related diseases such as PE [23]. Placental villi from pre-eclamptic and normal pregnancy patients of close gestational stages were analyzed by immunohistochemistry (Fig 7A) and were dissected and subjected to western blot analysis (Fig 7B). Immunohistochemistry showed weak positive signals for MSX2 in the EVTs from PE placentas (Fig 7Ab) compared with the strong MSX2 staining in the EVTs from normal placentas (Fig 7Aa). Western blotting(Fig 7B) and statistical analyses o(Fig 7C) indicated that MSX2 was expressed at significantly lower levels in the placental villi from pre-eclamptic patients than in those from control placentas (n = 8).

Bottom Line: Furthermore, we found that the in vitro invasive ability of HTR8/SVneo cells was enhanced by exogenous overexpression of MSX2, and that this effect was accompanied by increased protein expression of matrix metalloproteinase-2 (MMP-2), vimentin, and β-catenin.Meanwhile, overexpression of MSX2 and treatment with the MSX2-specific siRNA resulted in decreased and increased E-cadherin expression, respectively, in JEG-3 cells.Collectively, our results suggest that MSX2 may induce human trophoblast cell invasion, and dysregulation of MSX2 expression may be associated with pre-eclampsia.

View Article: PubMed Central - PubMed

Affiliation: Laboratory Animal Center, Chongqing Medical University, Chongqing, China.

ABSTRACT
Normal implantation depends on appropriate trophoblast growth and invasion. Inadequate trophoblast invasion results in pregnancy-related disorders, such as early miscarriage and pre-eclampsia, which are dangerous to both the mother and fetus. Msh Homeobox 2 (MSX2), a member of the MSX family of homeobox proteins, plays a significant role in the proliferation and differentiation of various cells and tissues, including ectodermal organs, teeth, and chondrocytes. Recently, MSX2 was found to play important roles in the invasion of cancer cells into adjacent tissues via the epithelial-mesenchymal transition (EMT). However, the role of MSX2 in trophoblastic invasion during placental development has yet to be explored. In the present study, we detected MSX2 expression in cytotrophoblast, syncytiotrophoblast, and extravillous cytotrophoblast cells of first or third trimester human placentas via immunohistochemistry analysis. Furthermore, we found that the in vitro invasive ability of HTR8/SVneo cells was enhanced by exogenous overexpression of MSX2, and that this effect was accompanied by increased protein expression of matrix metalloproteinase-2 (MMP-2), vimentin, and β-catenin. Conversely, treatment of HTR8/SVneo cells with MSX2-specific siRNAs resulted in decreased protein expression of MMP-2, vimentin, and β-catenin, and reduced invasion levels in a Matrigel invasion test. Notably, however, treatment with the MSX2 overexpression plasmid and the MSX2 siRNAs had no effect on the mRNA expression levels of β-catenin. Meanwhile, overexpression of MSX2 and treatment with the MSX2-specific siRNA resulted in decreased and increased E-cadherin expression, respectively, in JEG-3 cells. Lastly, the protein expression levels of MSX2 were significantly lower in human pre-eclamptic placental villi than in the matched control placentas. Collectively, our results suggest that MSX2 may induce human trophoblast cell invasion, and dysregulation of MSX2 expression may be associated with pre-eclampsia.

No MeSH data available.


Related in: MedlinePlus