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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

Effects of MSX2 overexpression or knockdown on proliferation and apoptosis of HTR8/SVneo cell.(A) MTT assay of HTR8/SVneo cell transfected with indicated vectors or siRNAs showed no significant difference on proliferation. (t-test). (B, D) Cell cycle progression was analyzed by flow cytometry at 24 h after transfection with the MSX2 overexpression vector or the MSX2 siRNA. Results were evaluated using MODFIT software. (C, E)Statistical bar graphs summarizing the results of three independent experiments in,respectively.(t-test,P>0.05; n = 3).
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pone.0153656.g004: Effects of MSX2 overexpression or knockdown on proliferation and apoptosis of HTR8/SVneo cell.(A) MTT assay of HTR8/SVneo cell transfected with indicated vectors or siRNAs showed no significant difference on proliferation. (t-test). (B, D) Cell cycle progression was analyzed by flow cytometry at 24 h after transfection with the MSX2 overexpression vector or the MSX2 siRNA. Results were evaluated using MODFIT software. (C, E)Statistical bar graphs summarizing the results of three independent experiments in,respectively.(t-test,P>0.05; n = 3).

Mentions: After Matrigel cell invasion assays, we further investigated the effects of MSX2 on trophoblast cell HTR8/SVneo proliferation and apoptosis. HTR8/SVneo cells transfected with the siRNA or the plasmids were subjected to MTT assay(Fig 4A) and subsequent flow cytometry analysis by PI staining(Fig 4B). Meanwhile, apoptosis levels were evaluated using a FITC-labeled annexin V-specific antibody (Fig 4D). Our results indicated that either MSX2 knock down or over expression had no obvious effects on cell proliferation or apoptosis compared with the control groups (P>0.05).


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)

Effects of MSX2 overexpression or knockdown on proliferation and apoptosis of HTR8/SVneo cell.(A) MTT assay of HTR8/SVneo cell transfected with indicated vectors or siRNAs showed no significant difference on proliferation. (t-test). (B, D) Cell cycle progression was analyzed by flow cytometry at 24 h after transfection with the MSX2 overexpression vector or the MSX2 siRNA. Results were evaluated using MODFIT software. (C, E)Statistical bar graphs summarizing the results of three independent experiments in,respectively.(t-test,P>0.05; n = 3).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4835074&req=5

pone.0153656.g004: Effects of MSX2 overexpression or knockdown on proliferation and apoptosis of HTR8/SVneo cell.(A) MTT assay of HTR8/SVneo cell transfected with indicated vectors or siRNAs showed no significant difference on proliferation. (t-test). (B, D) Cell cycle progression was analyzed by flow cytometry at 24 h after transfection with the MSX2 overexpression vector or the MSX2 siRNA. Results were evaluated using MODFIT software. (C, E)Statistical bar graphs summarizing the results of three independent experiments in,respectively.(t-test,P>0.05; n = 3).
Mentions: After Matrigel cell invasion assays, we further investigated the effects of MSX2 on trophoblast cell HTR8/SVneo proliferation and apoptosis. HTR8/SVneo cells transfected with the siRNA or the plasmids were subjected to MTT assay(Fig 4A) and subsequent flow cytometry analysis by PI staining(Fig 4B). Meanwhile, apoptosis levels were evaluated using a FITC-labeled annexin V-specific antibody (Fig 4D). Our results indicated that either MSX2 knock down or over expression had no obvious effects on cell proliferation or apoptosis compared with the control groups (P>0.05).

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