Korean J Obstet Gynecol.  2010 Oct;53(10):905-914. 10.5468/kjog.2010.53.10.905.

The effect of granulocyte-macrophage-stimulating factor on the gene expression in human trophoblast of early pregnancy

• Lee HY
• Kim SS
• Ahn CS
• Park YJ
• Jun JH
• Song H
• Moon HS
• Park EJ
• Hong SY
• Park WI

• KMID: 1453755
• DOI: http://doi.org/10.5468/kjog.2010.53.10.905

Abstract

OBJECTIVE
Granulocyte-macrophage colony-stimulating factor (GM-CSF) facilitates mammalian embryonic development and implantation. However, its biological function after implantation is not elucidated. The aim of this study is to assess the changes of gene expression by GM-CSF in human trophoblast obtained in early pregnancy.
METHODS
Human trophoblast obtained in early pregnancy was cultured with or without GM-CSF. The difference of gene expression was evaluated with microarray and selected genes were reevaluated with real-time reverse transcription-polymerase chain reaction (RT-PCR).
RESULTS
Microarray analysis revealed that the expressions of 468 genes were increased while those of 40 genes were decreased by GM-CSF. These genes were evaluated according to the known biologic pathways. The regulation of actin cytoskeleton and focal adhesion pathways were mostly influenced by GM-CSF. Annexin A2, thymosin-like 3, vimentin, myogenin, ACK1, and tensin1 genes were selected for real-time RT-PCR. The increased expressions of of vimentin and ACK1, and decreased expressions of tensin1 were confirmed by real-time RT-PCR.
CONCLUSION
GM-CSF activates focal adhesion pathway in human trophoblast by increasing the expression of vimentin and ACK1, and decreasing the expression of tensin1.

Keyword

Granulocyte-macrophage colony-stimulating factor; Trophoblast; Focal adhesion; Invasion

MeSH Terms

Actin Cytoskeleton
Annexin A2
Embryonic Development
Female
Focal Adhesions
Gene Expression
Granulocyte-Macrophage Colony-Stimulating Factor
Humans
Microarray Analysis
Myogenin
Pregnancy
Trophoblasts
Vimentin
Annexin A2
Granulocyte-Macrophage Colony-Stimulating Factor
Myogenin
Vimentin

Figure

• 

  Fig. 1 Diagram showing 28S and 18S peaks of each RNA sample. The ratio of 28S (left peak) and 18S (right peak) was calculated.

• 

  Fig. 2 Photoimage of microarray chip. Green dots represent genes their expression was increased by GM-SCF. Red dots represent genes their expression was decreased by GM-SCF. Linear plot of genes were found in sample E (A), G (B), and H (C). GM-CSF: granulocyte-macrophage colony-stimulating factor.

• 

  Fig. 3 Quantitative analysis of gene expressions by real time RT-PCR in human trophoblasts after GM-CSF treatment. *P<0.05 by Mann Whitney U test. GAPDH: glyceraldehyde-3-phosphatase dehydrogenase, ACK1: Cdc42-associated kinase, VIM: vimentin, MYOG: myogenin ANXA2: annexin A2, TMSL3: thymosin-like 3, TNS1: tensin1, PTPN2: protein tyrosine phosphatase, non-receptor type 2, GM-CSF: granulocyte-macrophage colony-stimulating factor.

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