Department of Biology, School of Medicine, University of Zagreb, Zagreb, Croatia.
Int J Exp Pathol. 2014 Aug;95(4):238-43. doi: 10.1111/iep.12088. Epub 2014 Jun 19.
The DNA demethylating agent 5-azacytidine (5-azaC) has a teratogenic influence during rat development influencing both the embryo and the placenta. Our aim was to investigate its impact on early decidual cell proliferation before the formation of placenta. Thus, female Fischer rats received 5-azaC (5 mg/kg, i.p.) on the 2nd, 5th or 8th day of gestation and the decidual tissues were harvested on gestation day 9. They were then analysed immunohistochemically for expression of cell proliferation marker proliferating cell nuclear antigen (PCNA) in decidual cells and for global DNA methylation using the coupled restriction enzyme digestion, random amplification and pyrosequencing assays. We found that 5-azaC administered on the 5th and 8th (but not on 2nd) day of gestation led to increased PCNA expression in decidual cells compared with untreated controls. No significant changes in DNA methylation were detected, with either method, in any of the treated rat groups compared with untreated controls. Thus, we conclude that 5-azaC can stimulate decidual cell proliferation without simultaneously changing global DNA methylation level in treated cells.
DNA 去甲基化剂 5-氮杂胞苷（5-azaC）在大鼠发育过程中具有致畸作用，影响胚胎和胎盘。我们的目的是研究其在胎盘形成前对早期蜕膜细胞增殖的影响。因此，雌性 Fischer 大鼠在妊娠第 2、5 或 8 天接受 5-azaC（5mg/kg，腹腔注射），并在妊娠第 9 天采集蜕膜组织。然后，使用偶联的限制性内切酶消化、随机扩增和焦磷酸测序检测蜕膜细胞中细胞增殖标志物增殖细胞核抗原（PCNA）的表达和整体 DNA 甲基化情况。我们发现，与未处理的对照组相比，在妊娠第 5 天和第 8 天（但不是第 2 天）给予 5-azaC 会导致蜕膜细胞中 PCNA 表达增加。与未处理的对照组相比，两种方法在任何处理组的大鼠中均未检测到 DNA 甲基化水平的显著变化。因此，我们得出结论，5-azaC 可以刺激蜕膜细胞增殖，而不会同时改变处理细胞中的整体 DNA 甲基化水平。
