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1.
Adv Exp Med Biol ; 1287: 47-57, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33034025

RESUMO

The human endometrium is a unique, highly dynamic tissue that undergoes cyclic changes of cell proliferation, differentiation, and death. Endometrial cancer is the most common malignancy among women in developed countries. Importantly, the incidence of endometrial cancer is rising in high-income countries. Currently histological classification is used for subtyping of endometrial cancer, while ongoing research is evaluating markers for more accurate molecular classification. Evolutionary conserved Notch signaling pathway regulates diverse cellular processes such as proliferation, differentiation, and cell invasion. Accumulating evidence links aberrant Notch signaling with diseases such as hyperplasia and endometrial cancer. This chapter summarizes the current state of Notch signaling investigations in the endometrium, endometriosis, and endometrial cancer.


Assuntos
Neoplasias do Endométrio , Endometriose , Receptores Notch/metabolismo , Neoplasias do Endométrio/metabolismo , Neoplasias do Endométrio/patologia , Endometriose/metabolismo , Endometriose/patologia , Endométrio/metabolismo , Endométrio/patologia , Feminino , Humanos , Transdução de Sinais
2.
Anim Sci J ; 91(1): e13456, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32926548

RESUMO

In this study, we investigated whether bovine oviducts and endometria produce anti-Müllerian hormone (AMH) (for paracrine and autocrine signaling). Reverse transcription-polymerase chain reaction and western blotting detected AMH expression in oviductal and endometrial specimens. Immunohistochemistry revealed robust AMH expression in the ampulla and isthmus epithelia, and the glandular and luminal endometrial epithelia (caruncular endometria). AMH mRNA (measured by real-time PCR) and protein expression in these layers did not significantly differ among estrous phases in adult Japanese Black (JB) heifers (p > .1). Furthermore, the expression in these layers also did not differ among Holstein cows (93.8 ± 5.8 months old), JB heifers (25.5 ± 0.4 months old), and JB cows (97.9 ± 7.9 months old). We also compared AMH concentrations in the oviduct and uterine horn fluids among the three groups (measured by immunoassays). Interestingly, the AMH concentration in the oviduct fluid, but not in the uterine horn fluid, of Holstein cows was lower than those in JB heifers and cows (p < .05). Therefore, bovine oviducts and endometria express AMH and likely secrete it into the oviduct and uterine fluids.


Assuntos
Hormônio Antimülleriano/genética , Hormônio Antimülleriano/metabolismo , Endométrio/metabolismo , Células Epiteliais/metabolismo , Expressão Gênica , Oviductos/metabolismo , Animais , Bovinos , Endométrio/citologia , Ciclo Estral/metabolismo , Feminino , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Útero/metabolismo
3.
Medicine (Baltimore) ; 99(37): e22163, 2020 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-32925779

RESUMO

For frozen embryo transplantation patients who failed to use hormone replacement cycle (HRC) transplantation for 2 consecutive times, the third time of transplantation was divided into 2 groups: HRC and natural cycle (NC), and the pregnancy rate of the 2 groups, especially the clinical pregnancy rate, was compared.Retrospective study of 174 patients in the reproductive medicine center of an affiliated hospital of Shandong University of Traditional Chinese Medicine between January 2015 and September 2018.The 174 patients were all infertile with regular menstruation. They had undergone 2 consecutive failed cycles of endometrial preparation with hormone replacement therapy and prepare for the third frozen embryo transplantation.A third cycle of treatment was planned using either NC or HRC for endometrial preparation. All the embryos were obtained during the same oocyte retrieval cycle. Patients were divided into groups based on the method of endometrial preparation: 98 were classified as NC and 76 as HRC.The pregnancy outcomes for the 2 groups were compared. Confounding factors that may affect clinical pregnancy rates were analyzed.We found that on the day of endometrial transformation, estrogen levels and endometrial thickness in the NC group were significantly higher than those in the HRC group. There were no significant differences in the rates of biochemical pregnancy, clinical pregnancy, cumulative pregnancy, miscarriage, multiple pregnancy, ectopic pregnancy, or live birth between the 2 groups. It is concluded by binary regression analysis that the different endometrial preparation protocol have no significant effect on the CPR.NC is as effective as HRC after 2 previous cycles of HRC. Because this was a retrospective study design, selection bias is possible, although the baseline characteristics of the 2 groups of patients were matched.


Assuntos
Transferência Embrionária/métodos , Terapia de Reposição Hormonal/métodos , Resultado da Gravidez/epidemiologia , Taxa de Gravidez , Adulto , Implantação do Embrião/fisiologia , Endométrio/metabolismo , Estrogênios/sangue , Feminino , Humanos , Gravidez , Estudos Retrospectivos
4.
PLoS One ; 15(7): e0234086, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32658928

RESUMO

Based on microRNA (miR) microarray analysis, we previously found that miR22-5p expression is decreased in the mid-luteal endometrium of women with minimal/mild endometriosis. Bioinformatics analysis predicted that miR22-5p targets ten-eleven translocation (TET2) 3'-untranslated region. This study aimed to determine the regulation and roles of miR22-5p in the pathogenesis of minimal/mild endometriosis-associated infertility. MiR22-5p and TET2 expression in the mid-luteal endometrium from women with or without minimal/mild endometriosis was analyzed. After transfection with miR22-5p mimics or inhibitor, TET2 expression was analyzed by quantitative reverse transcription (RT-q) PCR, western blotting and immunohistochemistry. 5-Hydroxymethylcytosine was determined by immunofluorescence and dot blotting. Expression and promoter methylation of estrogen receptor 2 (ESR2) was measured by RT-qPCR and western blotting, and by bisulfite sequencing, respectively. We first established that miR22-5p expression decreased and TET2 expression increased in minimal/mild endometriosis during implantation window. TET2 was found to be a direct target of miR22-5p. MiR22-5p regulated the expression of ESR2, but did not directly affect methylation of its promoter region (-197/+359). Our results suggest that an imbalance in miR22-5p expression in the mid-luteal endometrium may be involved in minimal/mild endometriosis-associated infertility.


Assuntos
Proteínas de Ligação a DNA/biossíntese , Implantação do Embrião , Endometriose/complicações , Endométrio/metabolismo , Receptor beta de Estrogênio/biossíntese , Infertilidade Feminina/etiologia , Fase Luteal/fisiologia , MicroRNAs/genética , Proteínas Proto-Oncogênicas/biossíntese , Regiões 3' não Traduzidas/genética , 5-Metilcitosina/análogos & derivados , 5-Metilcitosina/análise , Adulto , Células Cultivadas , Metilação de DNA , Proteínas de Ligação a DNA/genética , Receptor beta de Estrogênio/genética , Feminino , Regulação da Expressão Gênica , Genes Reporter , Humanos , Infertilidade Feminina/genética , MicroRNAs/antagonistas & inibidores , MicroRNAs/biossíntese , Regiões Promotoras Genéticas/genética , Transporte Proteico/genética , Proteínas Proto-Oncogênicas/genética , Células Estromais/metabolismo , Adulto Jovem
5.
Fertil Steril ; 114(2): 223-232, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32641214

RESUMO

OBJECTIVE: To determine the susceptibility of the endometrium to infection by-and thereby potential damage from-SARS-CoV-2. DESIGN: Analysis of SARS-Cov-2 infection-related gene expression from endometrial transcriptomic data sets. SETTING: Infertility research department affiliated with a public hospital. PATIENT(S): Gene expression data from five studies in 112 patients with normal endometrium collected throughout the menstrual cycle. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Gene expression and correlation between viral infectivity genes and age throughout the menstrual cycle. RESULT(S): Gene expression was high for TMPRSS4, CTSL, CTSB, FURIN, MX1, and BSG; medium for TMPRSS2; and low for ACE2. ACE2, TMPRSS4, CTSB, CTSL, and MX1 expression increased toward the window of implantation. TMPRSS4 expression was positively correlated with ACE2, CTSB, CTSL, MX1, and FURIN during several cycle phases; TMPRSS2 was not statistically significantly altered across the cycle. ACE2, TMPRSS4, CTSB, CTSL, BSG, and MX1 expression increased with age, especially in early phases of the cycle. CONCLUSION(S): Endometrial tissue is likely safe from SARS-CoV-2 cell entry based on ACE2 and TMPRSS2 expression, but susceptibility increases with age. Further, TMPRSS4, along with BSG-mediated viral entry into cells, could imply a susceptible environment for SARS-CoV-2 entry via different mechanisms. Additional studies are warranted to determine the true risk of endometrial infection by SARS-CoV-2 and implications for fertility treatments.


Assuntos
Betacoronavirus/metabolismo , Infecções por Coronavirus/metabolismo , Endométrio/metabolismo , Endométrio/virologia , Regulação Viral da Expressão Gênica , Pneumonia Viral/metabolismo , Adulto , Fatores Etários , Betacoronavirus/genética , Infecções por Coronavirus/genética , Feminino , Humanos , Ciclo Menstrual , Pessoa de Meia-Idade , Pandemias , Peptidil Dipeptidase A/biossíntese , Peptidil Dipeptidase A/genética , Pneumonia Viral/genética , Medição de Risco/métodos , Internalização do Vírus , Adulto Jovem
6.
Gene ; 757: 144926, 2020 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-32621951

RESUMO

Endometriosis is a frequent gynecologic disease in the world. CircRNAs can exert a crucial role in various diseases. Nevertheless, little is known about its roles in endometriosis. We investigated the involvement of circ_0061140 in endometriosis. Tissues from endometriosis women displayed an increased expression of circ_0061140. Then, we found loss of circ_0061140 significantly repressed ectopic endometrial cell proliferation, migration and invasion. Meanwhile,miR-140-3pcan demonstrate an important role in several cancers.Here, we reported miR-140-3p was reduced in ectopic endometrial cells and it acted as a target of circ_0061140. Moreover, miR-140-3p was able to reverse the effect of circ_0061140 on ectopic endometrial cells. Furthermore, Notch2 was predicted as a putative target of miR-140-3p. A positive correlation between circ_0061140 and Notch2 was indicated. miR-140-3p and Notch2 were operated as downstream effectors in the circ_0061140 mediated signaling in endometriosis. Decrease of circ_0061140 could depress endometriosis progression through modulating miR-140-3p and Notch2.


Assuntos
Movimento Celular , Proliferação de Células , Endometriose/genética , RNA Circular/genética , Células Cultivadas , Regulação para Baixo , Endometriose/metabolismo , Endometriose/patologia , Endométrio/metabolismo , Endométrio/patologia , Feminino , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Circular/metabolismo , Receptor Notch2/genética , Receptor Notch2/metabolismo
7.
Proc Natl Acad Sci U S A ; 117(25): 14532-14542, 2020 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-32513733

RESUMO

Implantation is initiated when an embryo attaches to the uterine luminal epithelium and subsequently penetrates into the underlying stroma to firmly embed in the endometrium. These events are followed by the formation of an extensive vascular network in the stroma that supports embryonic growth and ensures successful implantation. Interestingly, in many mammalian species, these processes of early pregnancy occur in a hypoxic environment. However, the mechanisms underlying maternal adaptation to hypoxia during early pregnancy remain unclear. In this study, using a knockout mouse model, we show that the transcription factor hypoxia-inducible factor 2 alpha (Hif2α), which is induced in subluminal stromal cells at the time of implantation, plays a crucial role during early pregnancy. Indeed, when preimplantation endometrial stromal cells are exposed to hypoxic conditions in vitro, we observed a striking enhancement in HIF2α expression. Further studies revealed that HIF2α regulates the expression of several metabolic and protein trafficking factors, including RAB27B, at the onset of implantation. RAB27B is a member of the Rab family of GTPases that allows controlled release of secretory granules. These granules are involved in trafficking MMP-9 from the stroma to the epithelium to promote luminal epithelial remodeling during embryo invasion. As pregnancy progresses, the HIF2α-RAB27B pathway additionally mediates crosstalk between stromal and endothelial cells via VEGF granules, developing the vascular network critical for establishing pregnancy. Collectively, our study provides insights into the intercellular communication mechanisms that operate during adaptation to hypoxia, which is essential for embryo implantation and establishment of pregnancy.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Hipóxia Celular/fisiologia , Implantação do Embrião/fisiologia , Vesículas Secretórias/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Comunicação Celular/fisiologia , Linhagem Celular , Embrião de Mamíferos , Endométrio/citologia , Endométrio/metabolismo , Feminino , Técnicas de Introdução de Genes , Humanos , Masculino , Camundongos , Camundongos Knockout , Gravidez , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Transdução de Sinais/fisiologia , Células Estromais , Proteínas rab de Ligação ao GTP/genética
8.
Nat Commun ; 11(1): 2717, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32483112

RESUMO

Somatic inactivating mutations of ARID1A, a SWI/SNF chromatin remodeling gene, are prevalent in human endometrium-related malignancies. To elucidate the mechanisms underlying how ARID1A deleterious mutation contributes to tumorigenesis, we establish genetically engineered murine models with Arid1a and/or Pten conditional deletion in the endometrium. Transcriptomic analyses on endometrial cancers and precursors derived from these mouse models show a close resemblance to human uterine endometrioid carcinomas. We identify transcriptional networks that are controlled by Arid1a and have an impact on endometrial tumor development. To verify findings from the murine models, we analyze ARID1AWT and ARID1AKO human endometrial epithelial cells. Using a system biology approach and functional studies, we demonstrate that ARID1A-deficiency lead to loss of TGF-ß tumor suppressive function and that inactivation of ARID1A/TGF-ß axis promotes migration and invasion of PTEN-deleted endometrial tumor cells. These findings provide molecular insights into how ARID1A inactivation accelerates endometrial tumor progression and dissemination, the major causes of cancer mortality.


Assuntos
Carcinogênese/genética , Carcinoma Endometrioide/genética , Reprogramação Celular/genética , Proteínas de Ligação a DNA/genética , Neoplasias do Endométrio/genética , Fatores de Transcrição/genética , Animais , Carcinogênese/metabolismo , Carcinoma Endometrioide/metabolismo , Carcinoma Endometrioide/patologia , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/metabolismo , Células Cultivadas , Proteínas de Ligação a DNA/metabolismo , Neoplasias do Endométrio/metabolismo , Neoplasias do Endométrio/patologia , Endométrio/citologia , Endométrio/metabolismo , Feminino , Perfilação da Expressão Gênica/métodos , Humanos , Camundongos da Linhagem 129 , Camundongos Endogâmicos BALB C , Camundongos Knockout , Camundongos Transgênicos , Mutação , Fatores de Transcrição/metabolismo
9.
PLoS Genet ; 16(6): e1008601, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32555663

RESUMO

Programmed cellular responses to cycling ovarian-derived steroid hormones are central to normal endometrial function. Abnormalities therein, as in the estrogen-dependent, progesterone-"resistant" disorder, endometriosis, predispose to infertility and poor pregnancy outcomes. The endometrial stromal fibroblast (eSF) is a master regulator of pregnancy success. However, the complex hormone-epigenome-transcriptome interplay in eSF by each individual steroid hormone, estradiol (E2) and/or progesterone (P4), under physiologic and pathophysiologic conditions, is poorly understood and was investigated herein. Genome-wide analysis in normal, early and late stage eutopic eSF revealed: i) In contrast to P4, E2 extensively affected the eSF DNA methylome and transcriptome. Importantly, E2 resulted in a more open versus closed chromatin, confirmed by histone modification analysis. Combined E2 with P4 affected a totally different landscape than E2 or P4 alone. ii) P4 responses were aberrant in early and late stage endometriosis, and mapping differentially methylated CpG sites with progesterone receptor targets from the literature revealed different but not decreased P4-targets, leading to question the P4-"resistant" phenotype in endometriosis. Interestingly, an aberrant E2-response was noted in eSF from endometriosis women; iii) Steroid hormones affected specific genomic contexts and locations, significantly enriching enhancers and intergenic regions and minimally involving proximal promoters and CpG islands, regardless of hormone type and eSF disease state. iv) In eSF from women with endometriosis, aberrant hormone-induced methylation signatures were mainly due to existing DNA methylation marks prior to hormone treatments and involved known endometriosis genes and pathways. v) Distinct DNA methylation and transcriptomic signatures revealed early and late stage endometriosis comprise unique disease subtypes. Taken together, the data herein, for the first time, provide significant insight into the hormone-epigenome-transcriptome interplay of each steroid hormone in normal eSF, and aberrant E2 response, distinct disease subtypes, and pre-existing epigenetic aberrancies in the setting of endometriosis, provide mechanistic insights into how endometriosis affects endometrial function/dysfunction.


Assuntos
Metilação de DNA , Endometriose/genética , Epigênese Genética , Estradiol/metabolismo , Progesterona/metabolismo , Transcriptoma , Adulto , Cromatina/genética , Cromatina/metabolismo , Ilhas de CpG , Endometriose/metabolismo , Endométrio/efeitos dos fármacos , Endométrio/metabolismo , Estradiol/farmacologia , Feminino , Humanos , Progesterona/farmacologia
10.
Nature ; 580(7805): 640-646, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32350471

RESUMO

All normal somatic cells are thought to acquire mutations, but understanding of the rates, patterns, causes and consequences of somatic mutations in normal cells is limited. The uterine endometrium adopts multiple physiological states over a lifetime and is lined by a gland-forming epithelium1,2. Here, using whole-genome sequencing, we show that normal human endometrial glands are clonal cell populations with total mutation burdens that increase at about 29 base substitutions per year and that are many-fold lower than those of endometrial cancers. Normal endometrial glands frequently carry 'driver' mutations in cancer genes, the burden of which increases with age and decreases with parity. Cell clones with drivers often originate during the first decades of life and subsequently progressively colonize the epithelial lining of the endometrium. Our results show that mutational landscapes differ markedly between normal tissues-perhaps shaped by differences in their structure and physiology-and indicate that the procession of neoplastic change that leads to endometrial cancer is initiated early in life.


Assuntos
Análise Mutacional de DNA , Endométrio/citologia , Endométrio/metabolismo , Epitélio/metabolismo , Saúde , Mutação , Adulto , Idade de Início , Idoso , Idoso de 80 Anos ou mais , Envelhecimento/genética , Carcinogênese/genética , Células Clonais/citologia , Neoplasias do Endométrio/genética , Endométrio/patologia , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Epitélio/patologia , Feminino , Humanos , Pessoa de Meia-Idade , Paridade/genética , Fatores de Tempo , Adulto Jovem
11.
Adv Exp Med Biol ; 1242: 1-12, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32406025

RESUMO

Intrauterine development of the uterus is promoted by the hormonal influence of the maternal steroid sex hormones on the female fetus. The cyclic activity of the endometrium starts at puberty, at menarche, and is controlled by the pituitary hormones (FSH and LH) and steroid ovarian hormones, the latter acting on the target tissue-the endometrium. The proliferative and secretory cyclic changes of the endometrium prepare the uterus for implantation of a fertilized ovum. Ovarian failure to secrete steroid hormones results in the menopausal gradual atrophy of the endometrium.


Assuntos
Hormônios Esteroides Gonadais/metabolismo , Útero/fisiologia , Endométrio/metabolismo , Feminino , Humanos , Ovário/metabolismo
12.
PLoS One ; 15(4): e0230044, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32236143

RESUMO

LEFTY2 (endometrial bleeding associated factor; EBAF or LEFTYA), a cytokine released shortly before menstrual bleeding, is a negative regulator of cell proliferation and tumour growth. LEFTY2 down-regulates Na+/H+ exchanger activity with subsequent inhibition of glycolytic flux and lactate production in endometrial cancer cells. Glucose can be utilized not only for glycolysis but also for glycogen formation. Both glycolysis and glycogen formation require cellular glucose uptake which could be accomplished by the Na+ coupled glucose transporter-1 (SGLT1; SLC5A1). The present study therefore explored whether LEFTY2 modifies endometrial SGLT1 expression and activity as well as glycogen formation. Ishikawa and HEC1a cells were exposed to LEFTY2, SGLT1 and glycogen synthase (GYS1) transcript levels determined by qRT-PCR. SGLT1, GYS1 and phospho-GYS1 protein abundance was quantified by western blotting, cellular glucose uptake from 2-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)Amino)-2-Deoxyglucose (2-NBDG) uptake, and cellular glycogen content utilizing an enzymatic assay and subsequent colorimetry. As a result, a 48-hour treatment with LEFTY2 significantly increased SGLT1 and GYS1 transcript levels as well as SGLT1 and GYS1 protein abundance in both Ishikawa and HEC1a cells. 2-NBDG uptake and cellular glycogen content were upregulated significantly in Ishikawa (type 1) but not in type 2 endometrial HEC1a cells, although there was a tendency of increased 2-NBDG uptake. Further, none of the effects were seen in human benign endometrial cells (HESCs). Interestingly, in both Ishikawa and HEC1a cells, a co-treatment with TGF-ß reduced SGLT1, GYS and phospho-GYS protein levels, and thus reduced glycogen levels and again HEC1a cells had no significant change. In conclusion, LEFTY2 up-regulates expression and activity of the Na+ coupled glucose transporter SGLT1 and glycogen synthase GYS1 in a cell line specific manner. We further show the treatment with LEFTY2 fosters cellular glucose uptake and glycogen formation and TGF-ß can negate this effect in endometrial cancer cells.


Assuntos
Neoplasias do Endométrio/metabolismo , Endométrio/metabolismo , Glicogênio Sintase/metabolismo , Glicogênio/metabolismo , Fatores de Determinação Direita-Esquerda/fisiologia , Transportador 1 de Glucose-Sódio/metabolismo , Linhagem Celular Tumoral , Feminino , Glucose/metabolismo , Humanos , Sódio/metabolismo
13.
PLoS One ; 15(4): e0231594, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32315343

RESUMO

Recurrence and poorly differentiated (grade 3 and above) and atypical cell type endometrial cancer (EC) have poor prognosis outcome. The mechanisms and characteristics of recurrence and distal metastasis of EC remain unclear. The extracellular matrix (ECM) of the reproductive tract in women undergoes extensive structural remodelling changes every month. Altered ECMs surrounding cells were believed to play crucial roles in a cancer progression. To decipher the associations between ECM and EC development, we generated a PAN-ECM Data list of 1516 genes including ECM molecules (ECMs), synthetic and degradation enzymes for ECMs, ECM receptors, and soluble molecules that regulate ECM and used RNA-Seq data from The Cancer Genome Atlas (TCGA) for the studies. The alterations of PAN-ECM genes by comparing the RNA-Seq expressions profiles of EC samples which have been grouped as tumorigenesis and metastasis group based on their pathological grading were identified. Differential analyses including functional enrichment, co-expression network, and molecular network analysis were carried out to identify the specific PAN-ECM genes that may involve in the progression of EC. Eight hundred and thirty-one and 241 PAN-ECM genes were significantly involved in tumorigenesis (p-value <1.571e-15) and metastasis (p-value <2.2e-16), respectively, whereas 140 genes were in the intersection of tumorigenesis and metastasis. Interestingly, 92 of the 140 intersecting PAN-ECM genes showed contrasting fold changes between the tumorigenesis and metastasis datasets. Enrichment analysis for the contrast PAN-ECM genes indicated pathways such as GP6 signaling, ILK signaling, and interleukin (IL)-8 signaling pathways were activated in metastasis but inhibited in tumorigenesis. The significantly activated ECM and ECM associated genes in GP6 signaling, ILK signaling, and interleukin (IL)-8 signaling pathways may play crucial roles in metastasis of EC. Our study provides a better understanding of the etiology and the progression of EC.


Assuntos
Carcinogênese/genética , Neoplasias do Endométrio/genética , Matriz Extracelular/genética , Proteínas de Neoplasias/genética , Biologia Computacional , Progressão da Doença , Neoplasias do Endométrio/patologia , Endométrio/metabolismo , Endométrio/patologia , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Metástase Neoplásica , RNA-Seq , Receptores de Superfície Celular/genética , Transdução de Sinais/genética
14.
Am J Physiol Cell Physiol ; 319(2): C392-C401, 2020 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-32348176

RESUMO

This study was conducted to define the underlying molecular mechanism of tripartite motif (TRIM) 59-induced invasion of ectopic endometrial stromal cells in endometriosis. Primary endometriosis ectopic endometrial stromal cells and normal endometrial cells were isolated and purified. Western blot was used to detect the expression of TRIM59, protein phosphatase Mg2+/Mn2+-dependent 1A (PPM1A), smad2/3, and phosphorylated (p)-smad2/3. Lentiviral vector-mediated TRIM59 interference and overexpression were established. Cell Counting Kit-8 assay was used to detect cell proliferation, and the Transwell migration assay was used to detect cell invasion. Matrix metalloproteinase (MMP-2), MMP9, smad2/3, and p-smad2/3 expressions were also detected using Western blot analysis; degradation of PPM1A was verified to be through ubiquitination. We found that TRIM59 expression levels in the endometriosis group was significantly higher compared with the normal group (P < 0.05), whereas the expression levels of PPM1A in the endometriosis group were significantly lower (P < 0.05). Endometriosis did not alter smad2/3 (P > 0.05) expression. However, after activating smad2/3 by phosphorylation, the expression of p-smad2/3 in the endometriosis group was significantly higher compared with the normal group (P < 0.05). The content of PPM1A in the TRIM59 overexpression group was significantly lower than that in the control group (P < 0.001), whereas the content of PPM1A in the siTRIM59 group was significantly higher than that in the control group (P < 0.001). In addition, there were no significant differences in the mRNA levels of PPM1A among the five groups, indicating that TRIM59 affects the expression of PPM1A at the posttranslational level (P < 0.05). Overexpression of TRIM59 significantly promoted the ubiquitination of PPM1A. We conclude that TRIM59 inhibits PPM1A through ubiquitination and activates the transforming growth factor-ß/Smad pathway to promote the invasion of ectopic endometrial stromal cells in endometriosis.


Assuntos
Endometriose/genética , Endométrio/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/genética , Proteína Fosfatase 2C/genética , Fator de Crescimento Transformador beta/genética , Proteínas com Motivo Tripartido/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Expressão Ectópica do Gene/genética , Endometriose/patologia , Endométrio/patologia , Feminino , Humanos , Cultura Primária de Células , Transdução de Sinais/genética , Proteína Smad2/genética , Células Estromais/metabolismo , Células Estromais/patologia , Ubiquitinação/genética
15.
Anim Sci J ; 91(1): e13350, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32219980

RESUMO

This study investigated the effect of type-I interferon (IFN) on the expression of matrix metalloproteinases (MMPs) of the bovine endometrial stromal cells (BES) and epithelial cells (BEE). The cells were separated and purified from the caruncles and cultured in DMEM/F-12 containing 10% fetal bovine serum. Spheroids were generated by using ascorbate. Zymograms of the supernatant showed that BEE predominantly expressed MMP-9, whereas MMP-2 was expressed in BES and homo-spheroids. While MMPs expression was not detected in hetero-spheroids. Real-time quantitative PCR revealed that type-I IFN and P4 suppressed the gene expression of MMP-2 and MMP-9 in hetero-spheroids, respectively. On the other hand, gelatin zymography analysis of the supernatant showed that type-I IFN strongly promote the clearance of MMPs. While zymograms of the MMPs stocked in the hetero-spheroids were significantly reduced by type-I IFN. Phenylmethanesulfonyl fluoride and leupeptin (both are serine proteinase inhibitors) significantly repressed the clearance of MMP-2 and MMP-9 induced by type-I IFN. Moreover, collagen fibers in hetero-spheroids significantly decreased after the treatment with type-I IFN. In conclusion, it was suggested that type-I IFN participate in the tissue remodeling by regulation the clearance of MMPs.


Assuntos
Bovinos/metabolismo , Endométrio/citologia , Endométrio/metabolismo , Expressão Gênica/efeitos dos fármacos , Interferon Tipo I/farmacologia , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Células Estromais/metabolismo , Animais , Células Cultivadas , Feminino , Humanos , Gravidez
16.
Gene ; 743: 144614, 2020 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-32222532

RESUMO

Buffalo reproduction struggles with a high incidence of early embryonic mortality. Effective treatment and prevention strategies for this condition are not available due to lack of understanding of molecular pathways in early pregnancy of this species. In the present study, we have attempted to understand these molecular pathways by characterizing the endometrial transcriptomic profiles of pregnant buffalos during early pregnancy. For the transcriptome profiling, buffalo endometrial tissues of 29-36 days of pregnancy and of nonpregnant luteal phase were collected from the local slaughterhouse. We confirmed the status of pregnancy based on the crown vertebral length of the foetus. Total RNA was isolated and sequencing was performed using the Illumina nextseq platform. The raw reads were filtered and mapped to the Bos taurus UMD 3.1 reference genome assembly. An average of 24,597 genes was investigated for differential expression between the two groups. Transcriptome data identified a total of 450 differentially expressed genes (using a cut off value of log2 fold changes >2 and <-2) in early pregnancy in comparison to the nonpregnant group (Padj < 0.05). Among these, 270 genes were significantly upregulated and 180 genes were downregulated. The most impacted pathways were related to secretion, transport, ionic homeostasis, mitosis and negative regulation of viral processes. In conclusion, our study characterized a unique set of DEGs, during the early pregnancy of buffalo, which potentially modulate the endometrial environment to establish and maintain a successful pregnancy.


Assuntos
Búfalos/fisiologia , Endométrio/metabolismo , Prenhez/genética , RNA/metabolismo , Transcriptoma/fisiologia , Animais , Regulação para Baixo , Feminino , Gravidez , RNA/genética , RNA/isolamento & purificação , RNA-Seq , Regulação para Cima
17.
J Pharmacol Sci ; 143(1): 1-8, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32094083

RESUMO

OBJECTIVES: To investigated the metrorrhagia volume-reduction activity, anti-inflammatory activity and repair-promoting activity of Clinopodium chinense (Benth.) O. Kuntze. METHODS: An abnormal uterine bleeding (AUB) model was induced via oral administration of mifepristone and misoprostol to pregnant rats, which were treated with the total extract of C. chinense (TEC). After 7 days, the metrorrhagia volume was measured, the levels of TXB2, 6-keto-PGF1α, IL-6 and TNF-α were measured by ELISA, the pathological changes and micro vessel density (MVD) of the endometrium were evaluated using HE and immunofluorescence staining, and the expression of VEGF, MMP-2/9 and TGF-ß were assessed by Western blotting. Preliminary phytochemicals were screened and identified by UPLC-Q-TOF-MS. RESULTS: Eleven compounds in C. chinense were identified via comparison to standard substances. The results of animal experiment showed TEC could reduce metrorrhagia volume, alleviate pathological injury and increase MVD to promote recovery of the endometrium; TEC could also increase the levels of TXB2 and the expression of VEGF, TGF-ß, decrease the levels of IL-6, TNF-α and the expression of MMP-2/9. CONCLUSIONS: TEC showed beneficial effects on treating AUB by reducing metrorrhagia volume, inhibiting the inflammatory response and promoting the repair of the endometrium. Additionally, TEC also showed great haemostatic potential in AUB.


Assuntos
Lamiaceae/química , Fitoterapia , Extratos Vegetais/administração & dosagem , Hemorragia Uterina/tratamento farmacológico , 6-Cetoprostaglandina F1 alfa/metabolismo , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Endométrio/irrigação sanguínea , Endométrio/metabolismo , Endométrio/patologia , Feminino , Mediadores da Inflamação/metabolismo , Interleucina-6/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Extratos Vegetais/isolamento & purificação , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
18.
Endocrinology ; 161(4)2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-32099996

RESUMO

Endometriosis is an inflammatory disease that primarily affects women during their reproductive years, and since current hormonal therapies are of concern, new hormone-independent treatment regimens are needed. The orphan nuclear receptor 4A1 (NR4A1, Nur77) is expressed in patient-derived (stromal) endometriotic cells and also epithelial cell lines, and we observed that knockdown of NR4A1 in patient-derived ectopic endometrium-isolated ovarian endometrioma (ESECT)-7 and ESECT-40 cells decreased cell proliferation and induced apoptosis. Moreover, the treatment of these cells with bis-indole derived NR4A1 ligands 1,1-bis(3'-indolyl)-1-(p-hydroxyphenyl)methane (DIM-C-pPhOH) and its buttressed 3-chloro-5-methoxy analog (DIM-C-pPhOH-3-Cl-5-OCH3) inhibited cell growth and induced apoptosis and related genes. The compounds exhibit NR4A1 antagonist activities in both functional and transactivation assays whereas these effects were not observed in normal endometrial cells. We also observed that NR4A1 knockdown and treatment with NR4A1 antagonists decreased fibrosis, α-smooth muscle actin, and related pro-fibrotic genes in ESECT-7 and ESECT-40 cells, and similar results were observed in epithelial-derived endometriotic cell lines. Moreover, in an endometriosis mouse model with auto-transplantation and also in severe combined immune deficiency mice transplanted with human endometriotic cells treatment with 25 mg/kg/day DIM-C-pPhOH-3-Cl-5-OCH3 significantly inhibited growth and expansion of endometriotic lesions. Thus, bis-indole-derived NR4A1 ligands represent a novel class of drugs as nonhormonal therapy for endometriosis.


Assuntos
Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Endometriose/tratamento farmacológico , Endométrio/efeitos dos fármacos , Indóis/farmacologia , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/antagonistas & inibidores , Fenóis/farmacologia , Animais , Modelos Animais de Doenças , Endometriose/metabolismo , Endométrio/metabolismo , Feminino , Técnicas de Silenciamento de Genes , Indóis/uso terapêutico , Camundongos , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Fenóis/uso terapêutico
19.
Fertil Steril ; 113(3): 670-678.e1, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32061358

RESUMO

OBJECTIVE: To study the prostanoid profile of the endometria of patients with recurrent implantation failure (RIF), unexplained infertility (UIF), and recurrent miscarriages (RM), and to compare them with the endometria of healthy fertile controls. DESIGN: Prospective cohort study. SETTING: University hospital. PATIENT(S): Fifteen patients with RIF, 18 patients with UIF, 16 patients with RM, and 23 fertile controls were recruited. INTERVENTION(S): Endometrial samples were taken during the window of implantation. After tissue homogenization and extraction, analysis with ultra-performance liquid chromatography diode array detector electrospray ionisation tandem mass spectrometry was performed. MAIN OUTCOME MEASURES: Concentrations of prostaglandin (PG) D1, PGE1, PGF1α, 6-ketoPGF1α, PGD2, PGE2, PGF2α, 15-deoxy-Δ12,14-PGJ2, PGD3, PGE3, PGF3α, thromboxane B2, 13,14-dihydro-PGE1, 13,14-dihydro-PGF1α, 13,14-dihydro-PGF2α, 13,14-dihydro-15-keto-PGE1, 13,14-dihydro-15-keto-PGE2, and 13,14-dihydro-15-keto-PGF2α were assessed. RESULT(S): Comparison of the endometria of patients with UIF and the controls showed no statistically significant differences. When the endometria of patients with RIF were compared with the controls, thromboxane B2 (TXB2) was found significantly higher (843.1 pg/mg vs. 133.5 pg/mg). When the endometria of patients with RM were compared with controls, 13,14-dihydro-15-keto PGF2α and TXB2 were found significantly higher (3907.30 pg/mg vs. 17.80 pg/mg and 858.7 pg/mg vs. 133.5 pg/mg respectively). CONCLUSION(S): We identified increased endometrial presence of TXB2 in patients with RM and RIF, and 13,14-dihydro-15-keto PGF2α in patients with RM. Although common ground is observed for RM and RIF, prostanoids, on the other hand, might make their own contribution to endometrial receptivity as important as genes and proteins. Attempts to normalize the prostaglandin profile of the endometrium via enzymatic activity can open new therapeutic options.


Assuntos
Aborto Habitual/metabolismo , Implantação do Embrião/fisiologia , Endométrio/metabolismo , Infertilidade Feminina/classificação , Infertilidade Feminina/metabolismo , Prostaglandinas/metabolismo , Aborto Habitual/patologia , Adulto , Biópsia , Estudos de Casos e Controles , Estudos de Coortes , Endométrio/química , Endométrio/patologia , Feminino , Humanos , Infertilidade Feminina/diagnóstico , Infertilidade Feminina/fisiopatologia , Metaboloma , Gravidez , Prostaglandinas/análise
20.
BMC Genomics ; 21(1): 103, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32000663

RESUMO

BACKGROUND: Luman is a member of CREB3 (cAMP responsive element-binding) subfamily of the basic leucine-zipper (bZIP) transcription factors. It may play an important regulatory role during the decidualization process since Luman was highly expressed in the decidual cells. However, the exact molecular mechanisms of how Luman regulating decidualization is unknown. RESULTS: Using an in vitro model, we prove that Luman knockdown significantly affects the decidualization process of mice endometrial stromal cells (ESCs) as the expression of two decidual markers PRL8a2 and PRL3c1 were repressed. We employed massively parallel RNA sequencing (RNA-Seq) to understand the changes in the transcriptional landscape associated with knockdown of Luman in ESCs during in vitro decidualization. We found significant dysregulation of genes related to protein processing in the endoplasmic reticulum (ER). Several genes involved in decidualization including bone morphogenetic proteins (e.g. BMP1, BMP4, BMP8A, BMP2, and BMP8B), growth factor-related genes (e.g. VEGFB, FGF10, and FGFR2), and transcription factors (IF4E, IF4A2, WNT4, WNT9A, ETS1, NOTCH1, IRX1, IDB1, IDB2, and IDB3), show altered expression. We also found that the knockdown of Luman is associated with increased expression of cell cycle-related genes including cycA1, cycB1, cycB2, CDK1, CDK2, and PLPK1, which resulted in an increased proportion of ESCs in the G1 phase. Differentially expressed genes (DEGs) were highly enriched on ECM-receptor interaction signaling, endoplasmic reticulum protein processing, focal adhesion, and PI3K-Akt signaling pathways. CONCLUSIONS: Luman knockdown results in widespread gene dysregulation during decidualization of ESCs. Genes involved in protein processing in ER, bone morphogenetic protein, growth factor, and cell cycle progression were identified as particularly important for explaining the decidual deficiency observed in this in vitro model. Therefore, this study provides clues as to the underlying mechanisms that may expand our understanding of gene regulation during decidualization.


Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Decídua/metabolismo , Perfilação da Expressão Gênica/veterinária , Redes Reguladoras de Genes , Animais , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Endométrio/metabolismo , Feminino , Regulação da Expressão Gênica , Técnicas de Silenciamento de Genes , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Camundongos , Análise de Sequência de RNA/veterinária
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