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Journal of Assisted Reproduction and Genetics

, Volume 34, Issue 2, pp 245–252 | Cite as

DNA methyltransferase 3A promoter polymorphism is associated with the risk of human spontaneous abortion after assisted reproduction techniques and natural conception

  • Yudong Liu
  • Haiyan Zheng
  • Pingping Guo
  • Shuxian Feng
  • Xingyu Zhou
  • Desheng Ye
  • Xin Chen
  • Shiling Chen
Epigenetics

Abstract

Purpose

The aim of this study was to explore the association of the DNA-methyltransferase (DNMT)-3A and DNMT3B promoter polymorphisms with the risk of human spontaneous abortion after assisted reproduction techniques (ARTs) and natural conception.

Methods

We collected tissues from women who underwent abortion procedures: (a) chorionic villus samples (CVS) and muscle samples (MS) from spontaneous abortions conceived by ART and natural cycle (study group), n = 152; and (b) CVS and MS from normal early pregnancy and second trimester (control group), n = 155. The single-nucleotide polymorphism (SNP) −448A > G in the DNMT3A promoter region and −149C/T polymorphism of DNMT3B were determined by polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) and confirmed by sequencing.

Results

The allele frequency of −448A among pregnancy loss group and control group was 34.2 % vs. 16.5 %, respectively. Compared with GG carriers, the DNMT3A −448AA homozygotes had an about 16-fold increased risk of spontaneous abortion [odds ratio (OR) = 16.130, 95 % confidence interval (CI), 3.665–70.984], and AG heterozygotes had an OR of 2.027 (95 % CI, 1.247–3.293). However, the distribution of −448A > G in individuals derived from ART pregnancies was not statistically significantly compared with those derived from spontaneous pregnancies (P = 0.661). For DNMT3B, we observed genotype frequencies of 100 % (TT) in the study group and the control group.

Conclusions

The DNMT3A −448A > G polymorphism may be a novel functional SNP and contribute to its genetic susceptibility to spontaneous abortion in Chinese women, and ART may not affect the distribution of −448A > G in pregnancy loss and normal pregnancy. The observed TT genotype of DMNT3B suggests that this is the predominant genotype of this population. The findings provide new insights into the etiology of human spontaneous abortion.

Keywords

DNA methylation Spontaneous abortion DNA-methyltransferase Single-nucleotide polymorphisms Assisted reproductive technology 

Notes

Acknowledgments

The authors gratefully acknowledge the technical support and valuable suggestions of all members from the Research Center of Clinical Medicine of Nanfang Hospital, and the help of the staff from the Department of Gynecology and Obstetrics in Nanfang Hospital in sample collection.

Compliance with ethical standards

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments.

Funding

This study was funded by the National Natural Science Foundation of China (no. 81170574 and no. 31371517) and High-level Project Matching Foundation of Nanfang Hospital, Southern Medical University (G201206).

Conflict of Interest

The authors declare that they have no competing interests.

Supplementary material

10815_2016_837_Fig2_ESM.gif (37 kb)
Figure S1

PCR-based restriction fragment length polymorphism genotyping of DNMT3A (a) and DNMT3B (b). a The wild-type G allele consists of a TaaI restriction site that results in two bands (153, 94, and 87 bp; 87 and 94 bp cannot be distinguished by electrophoresis, being combined into one band), the variant A allele also produces two bands (247 and 87 bp). Lanes 35, 7, 9, and 12: GG wild-type; lanes 1, 2, 6, 8, 11, and 13: AG heterozygotes; and lanes 10: AA variant (lanes 16, samples from the study group; lanes 713, samples from the control group). b The variant T allele had an AvrII restriction site that results in two bands (207 and 173 bp), and the wild-type C allele lacked the AvrII restriction site and therefore produced a single 380-bp band. Lanes 116: TT variant (lanes 1–8, samples from the study group; lanes 916, samples from the control group). (GIF 37 kb)

10815_2016_837_MOESM1_ESM.tif (15.2 mb)
High Resolution Image (TIF 15550 kb)

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Yudong Liu
    • 1
  • Haiyan Zheng
    • 1
    • 2
  • Pingping Guo
    • 1
  • Shuxian Feng
    • 1
  • Xingyu Zhou
    • 1
  • Desheng Ye
    • 1
  • Xin Chen
    • 1
  • Shiling Chen
    • 1
  1. 1.Center for Reproductive Medicine, Department of Gynecology and Obstetrics, Nanfang HospitalSouthern Medical UniversityGuangzhouPeople’s Republic of China
  2. 2.Center of Reproductive MedicineThird Affiliated Hospital of Guangzhou Medical UniversityGuangzhouPeople’s Republic of China

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