title: Methylated oligonucleotide (MON)-induced promoter hypermethylation is associated with repression of CDH1 expression and contributes to the migration and invasion of human trophoblast cell lines.
abstract: DNA cytosine-5 methylation plays a vital role in regulating the expression of E-cadherin, which is encoded by the CDH1 gene. In this study, we characterised the DNA methylation and expression pattern of CDH1 in an extravillous trophoblast cell line (HTR-8/SVneo) and two trophoblast cell lines -- JEG-3 and JAR. Promoter hypermethylation with reduced E-cadherin expression in HTR-8/SVneo cells and promoter hypomethylation with increased E-cadherin expression in JEG-3 and JAR cells were observed. Demethylation treatment significantly restored E-cadherin expression, contributing to decreases in the motility and invasiveness of HTR-8/SVneo cells. Sense-methylated oligonucleotides (MONs) labelled with Cy5 and complementary to a region of the human CDH1 promoter were designed, with the cytosines in 5'-cytosine-phosphate-guanine-3' (CpG) dinucleotides being replaced by methylated cytosines. Following MON transfection into JEG-3 cells, the level of CDH1 promoter DNA methylation as well as cell motility and invasiveness were increased and gene expression was significantly repressed. Our results indicate that MON-mediated DNA methylation of the CDH1 promoter and subsequent alterations in gene expression may contribute to trophoblast motility and invasion, suggesting a potential method for controlling the biological function of trophoblasts in vitro through epigenetic modification.
authors.affiliation: Laboratory of Reproductive Biology, School of Public Health and Management, Chongqing Medical University, No.1 Yixueyuan Rd, Chongqing, 400016, P.R. China.
authors.forename: Xi
authors.initials: X
authors.lastname: Lan
authors.affiliation: School of Traditional Chinese Medicine, Chongqing Medical University, No.1 Yixueyuan Rd, Chongqing, 400016, P.R. China.
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Fu
authors.affiliation: Department of Obstetrics and Gynecology, The First Affiliated Hospital of Chongqing Medical University, No.1 Youyi Rd, Chongqing, 400016, P.R. China.
authors.forename: Zhuo-Ying
authors.initials: ZY
authors.lastname: Hu
authors.affiliation: Laboratory of Reproductive Biology, School of Public Health and Management, Chongqing Medical University, No.1 Yixueyuan Rd, Chongqing, 400016, P.R. China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Feng
authors.affiliation: Laboratory of Reproductive Biology, School of Public Health and Management, Chongqing Medical University, No.1 Yixueyuan Rd, Chongqing, 400016, P.R. China.
authors.forename: Xue-Qing
authors.initials: XQ
authors.lastname: Liu
authors.affiliation: Laboratory of Reproductive Biology, School of Public Health and Management, Chongqing Medical University, No.1 Yixueyuan Rd, Chongqing, 400016, P.R. China.
authors.forename: Xue
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Laboratory of Reproductive Biology, School of Public Health and Management, Chongqing Medical University, No.1 Yixueyuan Rd, Chongqing, 400016, P.R. China.
authors.forename: Xue-Mei
authors.initials: XM
authors.lastname: Chen
authors.affiliation: Laboratory of Reproductive Biology, School of Public Health and Management, Chongqing Medical University, No.1 Yixueyuan Rd, Chongqing, 400016, P.R. China.
authors.forename: Jun-Lin
authors.initials: JL
authors.lastname: He
authors.affiliation: Laboratory of Reproductive Biology, School of Public Health and Management, Chongqing Medical University, No.1 Yixueyuan Rd, Chongqing, 400016, P.R. China.
authors.forename: Ying-Xiong
authors.initials: YX
authors.lastname: Wang
authors.affiliation: Laboratory of Reproductive Biology, School of Public Health and Management, Chongqing Medical University, No.1 Yixueyuan Rd, Chongqing, 400016, P.R. China.
authors.forename: Yu-Bin
authors.initials: YB
authors.lastname: Ding
mesh_terms: D015703:Antigens, CD; D015820:Cadherins / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects*; D019175:DNA Methylation / Q000187:drug effects*; D005260:Female; D006801:Humans; D009841:Oligonucleotides / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D014327:Trophoblasts / Q000166:cytology* / Q000187:drug effects
chemical_list: D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; D009841:Oligonucleotides