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1.
Reprod Biomed Online ; 32(6): 597-613, 2016 Jun.
Article in English | MEDLINE | ID: mdl-27090967

ABSTRACT

Little consensus has been reached on the best protocol for endometrial preparation for frozen embryo transfer (FET). It is not known how, and to what extent, hormone supplementation in artificial cycles influences endometrial preparation for embryo implantation at a molecular level, especially in patients who have experienced recurrent implantation failure. Transcriptome analysis of 15 endometrial biopsy samples at the time of embryo implantation was used to compare two different endometrial preparation protocols, natural versus artificial cycles, for FET in women who have experienced recurrent implantation failure compared with fertile women. IPA and DAVID were used for functional analyses of differentially expressed genes. The TRANSFAC database was used to identify oestrogen and progesterone response elements upstream of differentially expressed genes. Cluster analysis demonstrated that natural cycles are associated with a better endometrial receptivity transcriptome than artificial cycles. Artificial cycles seemed to have a stronger negative effect on expression of genes and pathways crucial for endometrial receptivity, including ESR2, FSHR, LEP, and several interleukins and matrix metalloproteinases. Significant overrepresentation of oestrogen response elements among the genes with deteriorated expression in artificial cycles (P < 0.001) was found; progesterone response elements predominated in genes with amended expression with artificial cycles (P = 0.0052).


Subject(s)
Embryo Implantation/physiology , Embryo Transfer/methods , Endometrium/pathology , Adult , Biopsy , Cluster Analysis , Cryopreservation/methods , Estradiol/therapeutic use , Estrogens/metabolism , Female , Gene Expression Profiling , Gene Expression Regulation , Hormones/metabolism , Humans , Matrix Metalloproteinases/metabolism , Oligonucleotide Array Sequence Analysis , Pregnancy , Pregnancy Rate , Principal Component Analysis , Progesterone/metabolism , Recurrence , Transcriptome , Treatment Outcome
2.
PLoS One ; 8(7): e68907, 2013.
Article in English | MEDLINE | ID: mdl-23874806

ABSTRACT

BACKGROUND: Estrogen (E2) and progesterone (P4) are key players in the maturation of the human endometrium. The corresponding steroid hormone modulators, tamoxifen (TAM) and mifepristone (RU486) are widely used in breast cancer therapy and for contraception purposes, respectively. METHODOLOGY/PRINCIPAL FINDINGS: Gene expression profiling of the human endometrial Ishikawa cancer cell line treated with E2 and P4 for 3 h and 12 h, and TAM and RU486 for 12 h, was performed using RNA-sequencing. High levels of mRNA were detected for genes, including PSAP, ATP5G2, ATP5H, and GNB2L1 following E2 or P4 treatment. A total of 82 biomarkers for endometrial biology were identified among E2 induced genes, and 93 among P4 responsive genes. Identified biomarkers included: EZH2, MDK, MUC1, SLIT2, and IL6ST, which are genes previously associated with endometrial receptivity. Moreover, 98.8% and 98.6% of E2 and P4 responsive genes in Ishikawa cells, respectively, were also detected in two human mid-secretory endometrial biopsy samples. TAM treatment exhibited both antagonistic and agonistic effects of E2, and also regulated a subset of genes independently. The cell cycle regulator cyclin D1 (CCND1) showed significant up-regulation following treatment with TAM. RU486 did not appear to act as a pure antagonist of P4 and a functional analysis of RU486 response identified genes related to adhesion and apoptosis, including down-regulated genes associated with cell-cell contacts and adhesion as CTNND1, JUP, CDH2, IQGAP1, and COL2A1. CONCLUSIONS: Significant changes in gene expression by the Ishikawa cell line were detected after treatments with E2, P4, TAM, and RU486. These transcriptome data provide valuable insight into potential biomarkers related to endometrial receptivity, and also facilitate an understanding of the molecular changes that take place in the endometrium in the early stages of breast cancer treatment and contraception usage.


Subject(s)
Endometrial Neoplasms/genetics , Estrogens/pharmacology , Mifepristone/pharmacology , Progesterone/pharmacology , Tamoxifen/pharmacology , Transcriptome/genetics , Cell Line, Tumor , Female , GTP-Binding Proteins/genetics , Gene Expression Profiling , Humans , Membrane Proteins/genetics , Mitochondrial Proteins/genetics , Neoplasm Proteins/genetics , Receptors for Activated C Kinase , Receptors, Cell Surface/genetics , Transcriptome/drug effects
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