Zheng Hai-Yan, Shi Xiao-Yun, Wang LE-LE, Wu Ya-Qin, Chen Shi-Ling, Zhang Lin
Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Nanfang Hospital, and.
Exp Ther Med. 2011 Jul;2(4):751-755. doi: 10.3892/etm.2011.261. Epub 2011 Apr 29.
Assisted reproductive technology (ART) including in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) have been shown to be associated with abnormal genomic imprinting, thus increasing the incidence of imprinting disorders such as Beckwith-Wiedemann syndrome (BWS) and Angelman syndrome (AS) in ART-conceived children. Furthermore, a recent study described abnormal DNA methylation in clinically normal children conceived by ART. However, data from different studies are conflicting or inconclusive. This study examined DNA methylation patterns of multiple imprinted genes in children born after ART to primarily evaluate the impact of ART on genomic imprinting. A total of 101 newborns conceived by ART (40 ICSI and 61 IVF) and 60 naturally conceived newborns were involved in our study. After obtaining the approval of the Institutional Ethics Committee, umbilical cord blood was collected from each infant. Genomic DNA was isolated from each blood sample and treated using sodium bisulfite. Subsequently, using methylation-specific PCR (MS-PCR), we analyzed six differentially methylated regions (DMRs) including KvDMR1, SNRPN, MEST, MEG3, TNDM and XIST. Meanwhile, information regarding twin pregnancies, gestational age, and birth weight of the neonates was documented. None of the cases presented with phenotypic abnormalities. Children conceived by ART were more likely to have low birth weight and to be born before term, compared with children conceived spontaneously. However, 7 months to 3 years of clinical follow-up showed that none of the children had clinical symptoms of any imprinting diseases. Furthermore, the MS-PCR results showed that all 161 children had normal DNA methylation patterns at six DMRs despite the different mode of conception. Our data did not indicate a higher risk of DNA-methylation defects in children born after ART. However, further studies using quantitative methods are needed to confirm these results.
包括体外受精（IVF）和卵胞浆内单精子注射（ICSI）在内的辅助生殖技术（ART）已被证明与异常的基因组印记有关，从而增加了ART受孕儿童中印记障碍的发生率，如贝克威思-维德曼综合征（BWS）和安吉尔曼综合征（AS）。此外，最近一项研究描述了ART受孕的临床正常儿童存在异常的DNA甲基化。然而，不同研究的数据相互矛盾或尚无定论。本研究检测了ART出生儿童多个印记基因的DNA甲基化模式，以主要评估ART对基因组印记的影响。我们的研究共纳入了101名ART受孕新生儿（40名ICSI和61名IVF）和60名自然受孕新生儿。在获得机构伦理委员会的批准后，从每个婴儿采集脐带血。从每个血样中分离基因组DNA并用亚硫酸氢钠处理。随后，使用甲基化特异性PCR（MS-PCR），我们分析了六个差异甲基化区域（DMR），包括KvDMR1、SNRPN、MEST、MEG3、TNDM和XIST。同时，记录了关于双胎妊娠、胎龄和新生儿出生体重的信息。所有病例均未出现表型异常。与自然受孕儿童相比，ART受孕儿童更有可能出生体重低且早产。然而，7个月至3年的临床随访表明，所有儿童均无任何印记疾病的临床症状。此外，MS-PCR结果显示，尽管受孕方式不同，但所有161名儿童在六个DMR处的DNA甲基化模式均正常。我们的数据并未表明ART出生儿童存在更高的DNA甲基化缺陷风险。然而，需要进一步使用定量方法的研究来证实这些结果。
