5-aza-2′-deoxycytidine leads to reduced embryo implantation and reduced expression of dna methyltransferases and essential endometrial genes5-aza-2u2032脱氧胞苷导致减少胚胎植入和dna甲基转移酶和必要的子宫内膜癌基因的表达减少.pdfVIP

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5-aza-2′-deoxycytidine leads to reduced embryo implantation and reduced expression of dna methyltransferases and essential endometrial genes5-aza-2u2032脱氧胞苷导致减少胚胎植入和dna甲基转移酶和必要的子宫内膜癌基因的表达减少.pdf

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5-aza-2′-deoxycytidine leads to reduced embryo implantation and reduced expression of dna methyltransferases and essential endometrial genes5-aza-2u2032脱氧胞苷导致减少胚胎植入和dna甲基转移酶和必要的子宫内膜癌基因的表达减少

5-Aza-29-deoxycytidine Leads to Reduced Embryo Implantation and Reduced Expression of DNA Methyltransferases and Essential Endometrial Genes Yu-Bin Ding, Chun-Lan Long, Xue-Qing Liu, Xue-Mei Chen, Liang-Rui Guo, Yin-Yin Xia, Jun-Lin He*, Ying- Xiong Wang* Department of Reproductive Biology, Chongqing Medical University, Chongqing, People’s Republic of China Abstract Background: The DNA demethylating agent 5-aza-2 9-deoxycytidine (5-aza-CdR) incorporates into DNA and decreases DNA methylation, sparking interest in its use as a potential therapeutic agent. We aimed to determine the effects of maternal 5- aza-CdR treatment on embryo implantation in the mouse and to evaluate whether these effects are associated with decreased levels of DNA methyltransferases (Dnmts) and three genes (estrogen receptor a [Esr1], progesterone receptor [Pgr], and homeobox A10 [Hoxa10]) that are vital for control of endometrial changes during implantation. Methods and Principal Findings: Mice treated with 5-aza-CdR had a dose-dependent decrease in number of implantation sites, with defected endometrial decidualization and stromal cell proliferation. Western blot analysis on pseudo-pregnant day 3 (PD3) showed that 0.1 mg/kg 5-aza-CdR significantly repressed Dnmt3a protein level, and 0.5 mg/kg 5-aza-CdR significantly repressed Dnmt1, Dnmt3a, and Dnmt3b protein levels in the endometrium. On PD5, mice showed significantly decreased Dnmt3a protein level with 0.1 mg/kg 5-aza-CdR, and significantly decreased Dnmt1 and Dnmt3a with 0.5 mg/kg 5-aza-CdR. Immunohistochemical staining showed that 5-aza-CdR repressed DNMT expression in a cell type–specific fashion within the uterus, including decreased expression of Dnmt1 in luminal and/or glandular epithelium and of Dnmt3a and Dnmt3b in stroma. Furthermore, the 59 flanking regions of the Esr1, Pgr, and Hoxa10 were hyp