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Lee, Kim, Ahn, Park, Jun, Song, Moon, Park, Hong, and Park: The effect of granulocyte-macrophage-stimulating factor on the gene expression in human trophoblast of early pregnancy
Original Article
Korean Journal of Obstetrics and Gynecology

Korean Journal of Obstetrics and Gynecology 2010; 53(10): 905-914.

Published online: 31 October 2010

DOI: https://doi.org/10.5468/kjog.2010.53.10.905

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

Hun Yul Lee, M.D.1, Sang Soo Kim, Ph.D.2, Chang Suk Ahn, M.S.2, Young Joo Park, B.S.2, Jin Hyun Jun, Ph.D.2, Haengsuk Song, Ph.D.3, Hye Sung Moon, M.D.4, Eun Joo Park, M.D.1, Seo You Hong, M.D.1, Won Il Park, M.D.1

1Department of Obstetrics and Gynecology, Eulji University School of Medicine, Daejeon, Korea.

2Life Science Institute, Eulji University School of Medicine, Daejeon, Korea.

3Laboratory of Molecular Oncology, Cheil General Hospital, Kwandong University College of Medicine, Seoul, Korea.

4Department of Obstetrics and Gynecology, Ewha Woman's University School of Medicine, Seoul, Korea.

Corresponding author (pwi3110@eulji.ac.kr)

Received 30 June 2010       Revised 3 September 2010       Accepted 17 September 2010

Copyright © 2010 Korean Society of Obstetrics and Gynecology

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.

Keywords: Granulocyte-macrophage colony-stimulating factor, Trophoblast, Focal adhesion, Invasion

Figures and Tables

Fig. 1
Diagram showing 28S and 18S peaks of each RNA sample. The ratio of 28S (left peak) and 18S (right peak) was calculated.
kjog-53-905-g001
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.
kjog-53-905-g002
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.
kjog-53-905-g003
Table 1
Primers for real-time RT-PCR
kjog-53-905-i001

RT-PCR: reverse transcription-polymerase chain reaction.

Table 2
Evaluation of mRNA quality by rRNA ratio in 6 matched samples
kjog-53-905-i002
Table 3
Function of genes up-regulated by GM-CSF in human trophoblast
kjog-53-905-i003

GM-CSF: granulocyte-macrophage colony-stimulating factor.

Table 4
Function of genes down-regulated by GM-CSF in human trophoblast
kjog-53-905-i004

GM-CSF: granulocyte-macrophage colony-stimulating factor.

Table 5
Selected 5 genes and their fold changes in microarray
kjog-53-905-i005

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