The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Reprod Sci. 2021 Apr;28(4):1133-1141. doi: 10.1007/s43032-020-00417-x. Epub 2021 Jan 29.
The aim of this study is to investigate the expression and DNA methylation status of the imprinted genes PEG10 and L3MBTL1 in the offspring of assisted reproductive technology (ART). The ART group consists of 30 cases of placenta and umbilical cord blood from ART full-term, uncomplicated singleton pregnancy progeny, and the normal control group consists of 30 cases of placenta and umbilical cord blood from natural full-term, uncomplicated singleton pregnancy progeny. The imprinted genes PEG10 and L3MBTL1 are analyzed, and the expression and methylation status of the two genes are detected using real-time quantitative polymerase chain reaction (QRT-PCR), immunohistochemistry (IHC), Western blotting (WB), and methylation-specific polymerase chain reaction (MSP). Compared with the normal control group, the PEG10 mRNA relative quantity (RQ) value in the placenta is 0.994 ± 0.458, with its RQ value up-regulated (P = 0.015). The PEG10 mRNA RQ value in the umbilical cord blood is 0.875 ± 0.452, with its RQ value up-regulated (P = 0.002). However, the L3MBTL1 mRNA RQ value in the placenta is 0.404 ± 0.234, with its RQ value down-regulated (P = 0.024). The L3MBTL1 mRNA RQ value in the umbilical cord blood is 0.337 ± 0.213, and there is no difference in the umbilical cord blood (P = 0.081). Compared with the normal control group, the expression of PEGl0 protein in the placenta of the ART progeny is up-regulated (P = 0.000), while the expression of L3MBTLl protein is down-regulated (P = 0.000). The methylation status of the PEGl0 promoter region in the placenta in the ART group is lower than that in the normal control group (P = 0.037), and that of the promoter region of the umbilical cord blood is lower than that of the natural pregnancy group (P = 0.032). The methylation status of the L3MBTLl promoter region is higher in the placenta than in the normal control group (P = 0.038), and there is no difference between the two groups in the umbilical cord blood (P = 0.301). In the ART group, the values of PEGl0 and L3MBTLl RQ in the placenta and the umbilical cord blood of the hypermethylated group are lower than in those of the hypomethylated group. ART may increase the risk of the abnormal expression of PEG10 and L3MBTL1 in offspring imprinted genes. The methylation of the promoter region may be the mechanism that regulates the expression of PEGl0 and L3MBTL1.
本研究旨在探讨辅助生殖技术（ART）子代印迹基因 PEG10 和 L3MBTL1 的表达和 DNA 甲基化状态。ART 组包括 30 例来自 ART 足月、无并发症的单胎妊娠后代的胎盘和脐血，正常对照组包括 30 例来自自然足月、无并发症的单胎妊娠后代的胎盘和脐血。分析印迹基因 PEG10 和 L3MBTL1，采用实时定量聚合酶链反应（QRT-PCR）、免疫组织化学（IHC）、Western blot（WB）和甲基化特异性聚合酶链反应（MSP）检测这两个基因的表达和甲基化状态。与正常对照组相比，胎盘 PEG10mRNA 相对定量（RQ）值为 0.994±0.458，其 RQ 值上调（P=0.015）。脐带血 PEG10mRNA RQ 值为 0.875±0.452，其 RQ 值上调（P=0.002）。然而，胎盘 L3MBTL1mRNA RQ 值为 0.404±0.234，其 RQ 值下调（P=0.024）。脐带血 L3MBTL1mRNA RQ 值为 0.337±0.213，且脐带血无差异（P=0.081）。与正常对照组相比，ART 后代胎盘中 PEG10 蛋白的表达上调（P=0.000），而 L3MBTL1 蛋白的表达下调（P=0.000）。ART 组胎盘 PEG10 启动子区的甲基化状态低于正常对照组（P=0.037），脐带血启动子区的甲基化状态低于自然妊娠组（P=0.032）。L3MBTL1 启动子区的甲基化状态高于正常对照组（P=0.038），两组脐带血无差异（P=0.301）。在 ART 组中，胎盘和脐带血高甲基化组 PEG10 和 L3MBTL1 的 RQ 值低于低甲基化组。ART 可能会增加印迹基因 PEG10 和 L3MBTL1 异常表达的风险。启动子区的甲基化可能是调节 PEG10 和 L3MBTL1 表达的机制。
