title: MicroRNA-212 Regulates the Expression of Olfactomedin 1 and C-Terminal Binding Protein 1 in Human Endometrial Epithelial Cells to Enhance Spheroid Attachment In Vitro.
abstract: Successful embryo implantation requires a synchronized dialogue between a competent blastocyst and the receptive endometrium, which occurs in a limited time period known as the "window of implantation." Recent studies suggested that down-regulation of olfactomedin 1 (OLFM1) in the endometrium and fallopian tube is associated with receptive endometrium and tubal ectopic pregnancy in humans. Interestingly, the human chorionic gonadotropin (hCG) induces miR-212 expression, which modulates OLFM1 and C-terminal binding protein 1 (CTBP1) expressions in mouse granulosa cells. Therefore, we hypothesized that embryo-derived hCG would increase miR-212 expression and down-regulate OLFM1 and CTBP1 expressions to favor embryo attachment onto the female reproductive tract. We found that hCG stimulated the expression of miR-212 and down-regulated OLFM1 but not CTBP1 mRNA in both human endometrial (Ishikawa) and fallopian (OE-E6/E7) epithelial cells. However, hCG suppressed the expression of OLFM1 and CTBP1 proteins in both cell lines. The 3'UTR of both OLFM1 and CTBP1 contained binding sites for miR-212. The miR-212 precursor suppressed luciferase expression, whereas the miR-212 inhibitor stimulated luciferase expression of the wild-type (WT)-OLFM1 and WT-CTBP1 reporter constructs. Furthermore, hCG (25 IU/ml) treatments stimulated trophoblastic (Jeg-3) spheroid (blastocyst surrogate) attachment onto Ishikawa and OE-E6/E7 cells. Transfection of miR-212 precursor increased Jeg-3 spheroid attachment onto Ishikawa cells and decreased OLFM1 and CTBP1 protein expressions, whereas the opposite occurred with miR-212 inhibitor. Taken together, hCG stimulated miR-212, which in turn down-regulated OLFM1 and CTBP1 expression in fallopian and endometrial epithelial cells to favor spheroid attachment.
authors.affiliation: Department of Obstetrics and Gynaecology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China Department of Veterinary Public Health and Pharmacology, Faculty of Veterinary Medicine and Animal Science, The University of Peradeniya, Peradeniya, Sri Lanka.
authors.forename: Kottawattage S A
authors.initials: KS
authors.lastname: Kottawatta
authors.affiliation: Department of Obstetrics and Gynaecology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China.
authors.forename: Kam-Hei
authors.initials: KH
authors.lastname: So
authors.affiliation: Department of Animal Science, Faculty of Agriculture, The University of Peradeniya, Peradeniya, Sri Lanka.
authors.forename: Suranga P
authors.initials: SP
authors.lastname: Kodithuwakku
authors.affiliation: Department of Obstetrics and Gynaecology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China Centre for Reproduction, Development and Growth, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China ShenZhen Key Laboratory of Fertility Regulation, The University of Hong Kong-Shenzhen Hospital, Futian District, Shenzhen, China.
authors.forename: Ernest H Y
authors.initials: EH
authors.lastname: Ng
authors.affiliation: Department of Obstetrics and Gynaecology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China Centre for Reproduction, Development and Growth, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China ShenZhen Key Laboratory of Fertility Regulation, The University of Hong Kong-Shenzhen Hospital, Futian District, Shenzhen, China.
authors.forename: William S B
authors.initials: WS
authors.lastname: Yeung
authors.affiliation: Department of Obstetrics and Gynaecology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China Centre for Reproduction, Development and Growth, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China ShenZhen Key Laboratory of Fertility Regulation, The University of Hong Kong-Shenzhen Hospital, Futian District, Shenzhen, China ckflee@hku.hk.
authors.forename: Kai-Fai
authors.initials: KF
authors.lastname: Lee
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D006063:Chorionic Gonadotropin; D004268:DNA-Binding Proteins / Q000378:metabolism*; D010064:Embryo Implantation; D004717:Endometrium / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005187:Fallopian Tubes / Q000378:metabolism; D005260:Female; D006023:Glycoproteins / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D018874:Spheroids, Cellular
chemical_list: D006063:Chorionic Gonadotropin; D004268:DNA-Binding Proteins; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; C551411:MIRN212 microRNA, human; D035683:MicroRNAs; C081451:olfactomedin; D000429:Alcohol Oxidoreductases; C096794:C-terminal binding protein
keywords: C-terminal binding protein 1; miR-212; microRNA; olfactomedin 1; spheroid attachment