Obstet Gynecol Sci.  2018 Sep;61(5):565-574. 10.5468/ogs.2018.61.5.565.

Differential expression and methylation of integrin subunit alpha 11 and thrombospondin in the amnion of preterm birth

• Yoo JY
• You YA
• Kwon EJ
• Park MH
• Shim S
• Kim YJ

• KMID: 2426021
• DOI: http://doi.org/10.5468/ogs.2018.61.5.565

Abstract

OBJECTIVE
This study aimed to investigate the association between preterm birth and epigenetic mechanisms in the amnion.
METHODS
We examined the association between differentially methylated regions (DMRs) and differentially expressed genes (DEG) using a cytosine-phosphate-guanine methylation array and whole-transcriptome sequencing from the amnion (preterm birth, n=5; full term, n=5). We enrolled 35 participants for mRNA expression analysis and pyrosequencing: 16 full-term and 19 preterm subjects. We compared the association of integrin subunit alpha 11 (ITGA11) and thrombospondin 2 (THBS2) gene methylation status with mRNA expression in the amnion.
RESULTS
In the preterm birth group, methylation of ITGA11 and THBS2 genes was significantly lower (ITGA11 gene: 60.30% vs. 73.16%, P < 0.05; THBS2 gene: 64.59% vs. 73.16%, P < 0.05), and the expression of the genes was significantly higher than that in the full-term group (ITGA11 gene: 14.20 vs. 1.57, P < 0.01; THBS2 gene: 1.18 vs. 10.34, P < 0.05).
CONCLUSION
Methylation of the ITGA11 and THBS2 genes in the amnion was associated with preterm birth. Thus, ITGA11 and THBS2 gene methylation status in the amnion may be valuable in explaining the mechanism underlying preterm birth.

Keyword

Preterm birth; Methylation; mRNA; ITGA11 protein; Thrombospondin 2; Gene expression

MeSH Terms

Amnion*
Epigenomics
Gene Expression
Methylation*
Parturition
Premature Birth*
RNA, Messenger
Thrombospondins*
RNA, Messenger
Thrombospondins

Figure

• 

  Fig. 1 Schematic representation of the human integrin subunit alpha 11 (A) and thrombospondin 2 (B) genes. Red line bar: pyrosequencing location; blue font: forward and reverse primer regions; violet font: sequencing primer region; green bar: real-time polymerase chain reaction location.

• 

  Fig. 2 Workflow for the sequence of data analyses with array-based technology and amnion samples. (A) DNA methylation values using an Illumina Infinitium II array. (B) mRNA expression detected using next-generation sequencing technology (Illumina NextSeq 500) and the amnion samples from preterm births.

• 

  Fig. 3 Methylation values of the integrin subunit alpha 11 (A) and thrombospondin 2 (B) genes in the amnion for the term and preterm births. mRNA expression levels of the integrin subunit alpha 11 (C) and thrombospondin 2 (D) genes in the amnion for the term and preterm births. The error bars represent standard deviations. ITGA11, integrin subunit alpha 11; THBS2, thrombospondin 2. a)P-values indicate significant differences between the term and preterm birth groups (non-parametric Wilcoxon rank sum test).

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