Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 23
Filter
2.
Endocrinology ; 165(1)2023 Nov 20.
Article in English | MEDLINE | ID: mdl-37972259

ABSTRACT

Interleukin 33 (IL-33) signaling regulates most of the key processes of pregnancy, including decidualization, trophoblast proliferation and invasion, vascular remodeling, and placental growth. Accordingly, dysregulation of IL-33, its membrane-bound receptor (ST2L, transducer of IL-33 signaling), and its soluble decoy receptor (sST2, inhibitor of IL-33 signaling) has been linked to a wide range of adverse pregnancy outcomes that are common in women with obesity and polycystic ovary syndrome, that is, conditions associated with hyperandrogenism, insulin resistance, and compensatory hyperinsulinemia. To reveal if androgens and insulin might modulate uteroplacental IL-33 signaling, we investigated the effect of dihydrotestosterone (DHT) and/or insulin on the expression of ST2L and sST2 (along with the activity of their promoter regions), IL-33 and sIL1RAP (heterodimerization partner of sST2), during in vitro decidualization of endometrial stromal cells from 9 healthy women. DHT and insulin markedly upregulated sST2 secretion, in addition to the upregulation of its messenger RNA (mRNA) expression, while the proximal ST2 promoter, from which the sST2 transcript originates, was upregulated by insulin, and in a synergistic manner by DHT and insulin combination treatment. On the other hand, sIL1RAP was slightly downregulated by insulin and IL-33 mRNA expression was not affected by any of the hormones, while ST2L mRNA expression and transcription from its promoter region (distal ST2 promoter) could not be detected or showed a negligibly low level. We hypothesize that high levels of androgens and insulin might lead to subfertility and pregnancy complications, at least partially, through the sST2-dependent downregulation of uteroplacental IL-33 signaling.


Subject(s)
Insulin , Interleukin-33 , Humans , Female , Pregnancy , Interleukin-33/genetics , Interleukin-33/metabolism , Interleukin-33/pharmacology , Insulin/pharmacology , Dihydrotestosterone/pharmacology , Interleukin-1 Receptor-Like 1 Protein/genetics , Interleukin-1 Receptor-Like 1 Protein/metabolism , Signal Transduction , Placenta/metabolism , Androgens/pharmacology , RNA, Messenger , Stromal Cells/metabolism
3.
BMJ Open ; 13(11): e073619, 2023 11 21.
Article in English | MEDLINE | ID: mdl-37989369

ABSTRACT

OBJECTIVE: Serum prokineticin-1 (s-PROK1) in the second and third trimester of pregnancy is positively correlated to preeclampsia, intrauterine growth restriction (IUGR) and preterm delivery. Women with polycystic ovary syndrome (PCOS) are prone to these adverse pregnancy outcomes. However, the contribution of PROK1 to the development of pregnancy complications and the effect of metformin and hyperandrogenism on s-PROK1 in PCOS have not been studied previously. DESIGN: This work is a post hoc analysis of two prospective, randomised, placebo-controlled trials. SETTING: Pregnant women with PCOS were included from 11 study centres in Norway. PARTICIPANTS: From 313 women, 264 participated in the present study after exclusions due to dropouts or insufficient serum samples. INTERVENTION: Women with PCOS were randomly administered with metformin or placebo, from first trimester to delivery. PRIMARY AND SECONDARY OUTCOME MEASURES: s-PROK1 was analysed using ELISA at gestational week 19 and related to pregnancy complications, fasting insulin levels, homoeostatic model assessment for insulin resistance (HOMA-IR), testosterone, or androstenedione levels, metformin use, PCOS phenotype and hyperandrogenism. RESULTS: Maternal s-PROK1 in the second trimester did not predict pregnancy-induced hypertension, pre-eclampsia or late miscarriage/preterm delivery in women with PCOS. However, s-PROK1 was lower in women who used metformin before inclusion, both in those randomised to metformin and to placebo, compared with those who did not. s-PROK1 was also lower in those who used metformin both at conception and during pregnancy compared with those who used metformin from inclusion or did not use metformin at all. s-PROK1 was lower in hyperandrogenic compared with normo-androgenic women with PCOS. CONCLUSIONS: Maternal s-PROK1 in the second trimester did not predict pregnancy complications in PCOS. Those who used metformin at conception and/or during pregnancy had lower s-PROK1. PCOS women with hyperandrogenism exhibited lower s-PROK1 compared with normo-adrogenic phenotypes. TRIAL REGISTRATION NUMBER: NCT03259919 and NCT00159536.


Subject(s)
Gastrointestinal Hormones , Hyperandrogenism , Metformin , Polycystic Ovary Syndrome , Pre-Eclampsia , Pregnancy Complications , Premature Birth , Vascular Endothelial Growth Factor, Endocrine-Gland-Derived , Infant, Newborn , Female , Pregnancy , Humans , Metformin/therapeutic use , Polycystic Ovary Syndrome/complications , Polycystic Ovary Syndrome/drug therapy , Hypoglycemic Agents/therapeutic use , Hyperandrogenism/chemically induced , Hyperandrogenism/complications , Hyperandrogenism/drug therapy , Prospective Studies , Pregnancy Complications/drug therapy , Pregnancy Complications/chemically induced , Pre-Eclampsia/drug therapy , Gastrointestinal Hormones/therapeutic use
4.
J Reprod Immunol ; 160: 104155, 2023 12.
Article in English | MEDLINE | ID: mdl-37801889

ABSTRACT

A recent meta-analysis revealed that patients with unexplained recurrent pregnancy loss (RPL) show higher insulin resistance compared to healthy controls. However, the etiology of RPL remains unknown. Prokineticin (PROK1), a pleiotropic uterine endometrial protein, is important for implantation and decidualization and is regulated by hypoxia and insulin. In this study, we investigated the decidualization status and the role of PROK1 in the decidua of patients with unexplained RPL showing insulin resistance. Thirty-two patients with unexplained RPL were included in this study. Following the diagnosis of a miscarriage, the decidua and villi of the patient were surgically collected. Fasting blood glucose and insulin levels were measured, and HOMA-ß was calculated. Using IHC and ELISA, the expression of IGFBP-1, PRL and PROK1 in the decidua and IGF-2 in the villi were analyzed in patients with euploid miscarriage with a high HOMA-ß index (n = 8) and compared to controls (euploid miscarriage with normal HOMA-ß: n = 12, aneuploid miscarriage with normal HOMA-ß: n = 12). The co-localization of PROK1 and IGFBP-1 was observed in the decidua by IHC. In the decidua of RPL patients with high HOMA-ß, the expression levels of IGFBP-1 and PRL were significantly lower, whereas the PROK1/IGFBP-1 ratio was significantly higher compared to that of the controls. IGF-2 expression in villi was significantly lower in RPL patients with high HOMA-ß. Impaired decidualization and excessive PROK1 production may have pathological implications in patients with unexplained RPL with insulin resistance, especially under the state of hyper insulin production.


Subject(s)
Abortion, Habitual , Gastrointestinal Hormones , Insulin Resistance , Vascular Endothelial Growth Factor, Endocrine-Gland-Derived , Pregnancy , Female , Humans , Decidua/pathology , Insulin-Like Growth Factor Binding Protein 1/metabolism , Insulin-Like Growth Factor II/metabolism , Abortion, Habitual/pathology , Insulin , Gastrointestinal Hormones/metabolism , Vascular Endothelial Growth Factor, Endocrine-Gland-Derived/metabolism
5.
Int J Mol Sci ; 24(4)2023 Feb 18.
Article in English | MEDLINE | ID: mdl-36835533

ABSTRACT

Recent studies suggest estradiol (E2)/natural progesterone (P) confers less breast cancer risk compared with conjugated equine estrogens (CEE)/synthetic progestogens. We investigate if differences in the regulation of breast cancer-related gene expression could provide some explanation. This study is a subset of a monocentric, 2-way, open observer-blinded, phase 4 randomized controlled trial on healthy postmenopausal women with climacteric symptoms (ClinicalTrials.gov; EUCTR-2005/001016-51). Study medication was two 28-day cycles of sequential hormone treatment with oral 0.625 mg CEE and 5 mg of oral medroxyprogesterone acetate (MPA) or 1.5 mg E2 as percutaneous gel/day with the addition of 200 mg oral micronized P. MPA and P were added days 15-28/cycle. Material from two core-needle breast biopsies in 15 women in each group was subject to quantitative PCR (Q-PCR). The primary endpoint was a change in breast carcinoma development gene expression. In the first eight consecutive women, RNA was extracted at baseline and after two months of treatment and subjected to microarray for 28856 genes and Ingenuity Pathways Analysis (IPA) to identify risk factor genes. Microarray analysis showed 3272 genes regulated with a fold-change of >±1.4. IPA showed 225 genes belonging to mammary-tumor development function: 198 for CEE/MPA vs. 34 for E2/P. Sixteen genes involved in mammary tumor inclination were subject to Q-PCR, inclining the CEE/MPA group towards an increased risk for breast carcinoma compared to the E2/P group at a very high significance level (p = 3.1 × 10-8, z-score 1.94). The combination of E2/P affected breast cancer-related genes much less than CEE/MPA.


Subject(s)
Medroxyprogesterone Acetate , Neoplasms , Humans , Female , Medroxyprogesterone Acetate/therapeutic use , Progesterone/adverse effects , Estrogens, Conjugated (USP)/pharmacology , Estradiol , Postmenopause , Estrogen Replacement Therapy/adverse effects , Risk Factors , Gene Expression , Neoplasms/drug therapy
6.
Oncoimmunology ; 11(1): 2109861, 2022.
Article in English | MEDLINE | ID: mdl-35979386

ABSTRACT

Tyrosine kinase inhibitors (TKIs) have dramatically improved the survival in chronic myeloid leukemia (CML), but residual disease typically persists even after prolonged treatment. Several lines of evidence suggest that TKIs administered to CML patients upregulate interferon γ (IFNγ) production, which may counteract the anti-tumorigenic effects of the therapy. We now show that activated T cell-conditioned medium (TCM) enhanced proliferation and counteracted imatinib-induced apoptosis of CML cells, and addition of a neutralizing anti-IFNγ antibody at least partially inhibited the anti-apoptotic effect. Likewise, recombinant IFNγ also reduced imatinib-induced apoptosis of CML cells. This anti-apoptotic effect of IFNγ was independent of alternative IFNγ signaling pathways, but could be notably diminished by STAT1-knockdown. Furthermore, IFNγ upregulated the expression of several anti-apoptotic proteins, including MCL1, PARP9, and PARP14, both in untreated and imatinib-treated primary human CD34+ CML stem/progenitor cells. Our results suggest that activated T cells in imatinib-treated CML patients can directly rescue CML cells from imatinib-induced apoptosis at least partially through the secretion of IFNγ, which exerts a rapid, STAT1-dependent anti-apoptotic effect potentially through the simultaneous upregulation of several key hematopoietic survival factors. These mechanisms may have a major clinical impact, when targeting residual leukemic stem/progenitor cells in CML.


Subject(s)
Interferon-gamma , Leukemia, Myelogenous, Chronic, BCR-ABL Positive , Antigens, CD34/metabolism , Antigens, CD34/pharmacology , Apoptosis , Cell Line, Tumor , Humans , Imatinib Mesylate/pharmacology , Imatinib Mesylate/therapeutic use , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/metabolism , Stem Cells/metabolism , Up-Regulation
7.
Cell Death Dis ; 12(10): 875, 2021 09 25.
Article in English | MEDLINE | ID: mdl-34564697

ABSTRACT

Tyrosine kinase inhibitor (TKI) treatment has dramatically improved the survival of chronic myeloid leukemia (CML) patients, but measurable residual disease typically persists. To more effectively eradicate leukemia cells, simultaneous targeting of BCR-ABL1 and additional CML-related survival proteins has been proposed. Notably, several highly specific myeloid cell leukemia 1 (MCL1) inhibitors have recently entered clinical trials for various hematologic malignancies, although not for CML, reflecting the insensitivity of CML cell lines to single MCL1 inhibition. Here, we show that combining TKI (imatinib, nilotinib, dasatinib, or asciminib) treatment with the small-molecule MCL1 inhibitor S63845 exerted strong synergistic antiviability and proapoptotic effects on CML lines and CD34+ stem/progenitor cells isolated from untreated CML patients in chronic phase. Using wild-type BCR-ABL1-harboring CML lines and their T315I-mutated sublines (generated by CRISPR/Cas9-mediated homologous recombination), we prove that the synergistic proapoptotic effect of the drug combination depended on TKI-mediated BCR-ABL1 inhibition, but not on TKI-related off-target mechanisms. Moreover, we demonstrate that colony formation of CML but not normal hematopoietic stem/progenitor cells became markedly reduced upon combination treatment compared to imatinib monotherapy. Our results suggest that dual targeting of MCL1 and BCR-ABL1 activity may efficiently eradicate residual CML cells without affecting normal hematopoietic stem/progenitors.


Subject(s)
Antineoplastic Agents/pharmacology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/pathology , Protein Kinase Inhibitors/pharmacology , Pyrimidines/pharmacology , Thiophenes/pharmacology , Antigens, CD34/metabolism , Antineoplastic Combined Chemotherapy Protocols , Apoptosis/drug effects , Caspase 3/metabolism , Caspase 7/metabolism , Cell Line, Tumor , Cell Survival/drug effects , Clone Cells , Drug Resistance, Neoplasm/drug effects , Drug Synergism , Hematopoietic Stem Cells/drug effects , Hematopoietic Stem Cells/metabolism , Humans , Imatinib Mesylate/administration & dosage , Imatinib Mesylate/pharmacology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/metabolism , Myeloid Cell Leukemia Sequence 1 Protein/antagonists & inhibitors , Myeloid Cell Leukemia Sequence 1 Protein/metabolism , Neoplasm Proteins/metabolism , Phosphate-Binding Proteins/metabolism , Poly(ADP-ribose) Polymerases/metabolism , Pore Forming Cytotoxic Proteins/metabolism , Pyroptosis/drug effects , Small Molecule Libraries/pharmacology , bcl-X Protein/metabolism
8.
J Cell Mol Med ; 25(20): 9523-9532, 2021 10.
Article in English | MEDLINE | ID: mdl-34463022

ABSTRACT

Finely tuned decidualization of endometrial stromal fibroblasts into decidual cells is crucial for successful implantation and a healthy pregnancy. Both insulin and androgens are known to modulate decidualization, however, their complex effect on this process has not been fully elucidated. As hyperinsulinemia and hyperandrogenism are associated in clinical conditions, we aimed to investigate the interaction between insulin and androgens on decidualization. Primary human endometrial stromal cells were decidualized in vitro in the presence of insulin and/or androgens (dihydrotestosterone (DHT), testosterone). Gene or protein expressions of decidualization markers were measured, and cells size characteristics were determined. Migration of decidualizing endometrial stromal cells and invasion of HTR-8/SVneo trophoblast spheroids were assessed. We found that insulin and androgens in combination enhanced the upregulation of several decidualization markers including prolactin, tissue factor, tissue inhibitor of matrix metalloproteinase 3 and connexin-43, and also interacted in modulating cell size characteristics resulting in enlarged decidualizing cells. However, insulin and DHT together restricted the migration of decidualizing cells and invasion of trophoblast spheroids. Our findings suggest that insulin and androgens interact to potentiate the process of decidualization. On the other hand, inhibited cell migration and trophoblast invasion might negatively impact the function of decidualizing endometrial stromal cells.


Subject(s)
Androgens/metabolism , Decidua/metabolism , Insulin/metabolism , Signal Transduction , Trophoblasts/metabolism , Androgens/pharmacology , Biomarkers , Cell Movement , Cells, Cultured , Coculture Techniques , Endometrium/cytology , Endometrium/metabolism , Female , Gap Junctions/metabolism , Gene Expression Regulation/drug effects , Humans , Immunophenotyping , Insulin/pharmacology , Pregnancy , Stromal Cells/metabolism
9.
Reprod Biol Endocrinol ; 18(1): 117, 2020 Nov 21.
Article in English | MEDLINE | ID: mdl-33218355

ABSTRACT

BACKGROUND: Solute carrier family 2 member 1 (SLC2A1; previously known as glucose transporter 1), is the most abundant glucose transporter in human endometrium and is up-regulated during decidualization, whereas high insulin may have a negative impact on this process. The present study aimed to investigate the effect of insulin on the expression of SLC2A1 and glucose uptake in decidualizing human endometrial stromal cells. METHODS: We induced in vitro decidualization of endometrial stromal cells obtained from regularly menstruating healthy non-obese women. The cells were treated with increasing concentrations of insulin, and the involvement of the transcription factor forkhead box O1 (FOXO1) was evaluated using a FOXO1 inhibitor. SLC2A1 mRNA levels were measured by Real-Time PCR and protein levels were evaluated by immunocytochemistry. Glucose uptake was estimated by an assay quantifying the cellular uptake of radioactive glucose. One-way ANOVA, Dunnett's multiple comparisons test and paired t-test were used to determine the statistical significance of the results. RESULTS: We found that insulin dose-dependently decreased SLC2A1 mRNA levels and decreased protein levels of SLC2A1 in decidualizing human endometrial stromal cells. Transcriptional inactivation of FOXO1 seems to explain at least partly the down-regulation of SLC2A1 by insulin. Glucose uptake increased upon decidualization, whereas insulin treatment resulted in a slight inhibition of the glucose uptake, although not significant for all insulin concentrations. CONCLUSIONS: These results indicate an impairment of decidualization by high concentrations of insulin. Future studies will determine the clinical significance of our results for endometrial function and decidualization in women with insulin resistance and hyperinsulinemia.


Subject(s)
Gene Expression/drug effects , Glucose Transporter Type 1/genetics , Glucose/metabolism , Insulin/pharmacology , Stromal Cells/drug effects , Adult , Cells, Cultured , Decidua/physiology , Down-Regulation/drug effects , Endometrium/cytology , Female , Glucose/pharmacokinetics , Glucose Transporter Type 1/metabolism , Humans , Hypoglycemic Agents/pharmacology , Immunohistochemistry , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Stromal Cells/metabolism , Young Adult
10.
J Cell Mol Med ; 24(5): 3242-3245, 2020 03.
Article in English | MEDLINE | ID: mdl-31991505

ABSTRACT

Prokineticin 1 (PROK1) is a key regulator of embryo implantation and placentation, and its dysregulation is associated with pregnancy complications, such as pre-eclampsia and foetal growth restriction. We have previously shown that insulin strongly enhances the expression of PROK1 in human decidualizing stromal cells. Here, we demonstrate that dihydrotestosterone (DHT), but not testosterone, potentiates insulin to up-regulate PROK1 in these cells. However, the androgens alone do not influence the expression of PROK1. Our findings suggest that insulin and androgens both are involved in the regulation of PROK1 that could have implications for normal and pathological pregnancies.


Subject(s)
Dihydrotestosterone/pharmacology , Endometrium/metabolism , Gastrointestinal Hormones/genetics , Insulin/genetics , Vascular Endothelial Growth Factor, Endocrine-Gland-Derived/genetics , Adolescent , Adult , Androgens/genetics , Biopsy , Cells, Cultured , Decidua/metabolism , Decidua/pathology , Embryo Implantation/genetics , Endometrium/drug effects , Endometrium/growth & development , Epithelial Cells/drug effects , Female , Gene Expression Regulation, Developmental/drug effects , Humans , Placentation/genetics , Pre-Eclampsia , Pregnancy , Pregnancy Complications/drug therapy , Pregnancy Complications/genetics , Pregnancy Complications/pathology , Primary Cell Culture , Signal Transduction/genetics , Stromal Cells/drug effects , Stromal Cells/pathology , Transcriptional Activation , Young Adult
11.
Biochem Biophys Res Commun ; 498(3): 502-508, 2018 04 06.
Article in English | MEDLINE | ID: mdl-29510136

ABSTRACT

B-cell CLL/lymphoma 6 (BCL6) is a transcriptional master regulator that can repress more than 1200 potential target genes. It exerts oncogenic effects through the inhibition of differentiation, DNA damage sensing and apoptosis in several human hematopoietic malignancies, including multiple myeloma (MM). The multifunctional cytokine interferon γ (IFNγ) exerts pro-apoptotic and anti-proliferative effects on MM cells in vitro, at least partially through the inhibition of the effects of interleukin 6 (IL6), one of the most important growth factor of MM and a strong inducer of BCL6 expression. However, IFNγ was also reported to directly upregulate BCL6 in several cell types. These observations prompted us to analyze the effect of IFNγ on BCL6 expression in MM cells. We discovered that among several myeloma growth/survival factors tested (including IL6, oncostatin M, insulin-like growth factor 1, tumor necrosis factor α and IFNα) IFNγ was the strongest inducer of BCL6 mRNA and protein expression in MM cell lines. IFNγ induced upregulation of BCL6 was dependent on the classical STAT1 signaling pathway, and affected both major BCL6 variants. Interestingly, although IFNα induced stronger STAT1 phosphorylation than IFNγ, it only slightly upregulated BCL6 in MM lines. We proved that IFNα induced BCL6 upregulation was limited by the concomitant activation of STAT5 signaling. We assume that BCL6 upregulation may represent a potentially pro-tumorigenic effect of IFNγ signaling in MM cells.


Subject(s)
Gene Expression Regulation, Neoplastic , Interferon-gamma/metabolism , Multiple Myeloma/genetics , Proto-Oncogene Proteins c-bcl-6/genetics , STAT1 Transcription Factor/metabolism , Up-Regulation , Cell Line, Tumor , Humans , Multiple Myeloma/metabolism , STAT5 Transcription Factor/metabolism , Signal Transduction , Tumor Suppressor Proteins/metabolism
12.
J Cell Mol Med ; 22(1): 163-172, 2018 01.
Article in English | MEDLINE | ID: mdl-28782224

ABSTRACT

Prokineticin 1 (PROK1), a hypoxia-regulated angiogenic factor, has emerged as a crucial regulator of embryo implantation and placentation. Dysregulation of PROK1 has been linked to recurrent pregnancy loss, pre-eclampsia, foetal growth restriction and preterm birth. These pregnancy complications are common in women with obesity and polycystic ovary syndrome, i.e. conditions associated with insulin resistance and compensatory hyperinsulinaemia. We investigated the effect of insulin on PROK1 expression during in vitro decidualization. Endometrial stromal cells were isolated from six healthy, regularly menstruating women and decidualized in vitro. Insulin induced a significant dose-dependent up-regulation of PROK1 on both mRNA and protein level in decidualizing endometrial stromal cells. This up-regulation was mediated by hypoxia-inducible factor 1-alpha (HIF1α) via the phosphatidylinositol 3-kinase (PI3K) pathway. Furthermore, we demonstrated that PROK1 did not affect the viability, but significantly inhibited the migration of endometrial stromal cells and the migratory and invasive capacity of trophoblast cell lines. This in vitro study provides new insights into the regulation of PROK1 by insulin in human decidualizing endometrial stromal cells, the action of PROK1 on migration of endometrial stromal cells, as well as migration and invasion of trophoblasts. We speculate that hyperinsulinaemia may be involved in the mechanisms by which PROK1 is linked to placenta-related pregnancy complications.


Subject(s)
Decidua/cytology , Decidua/metabolism , Gastrointestinal Hormones/genetics , Insulin/pharmacology , Vascular Endothelial Growth Factor, Endocrine-Gland-Derived/genetics , Adolescent , Adult , Cell Movement/drug effects , Cell Survival/drug effects , Choriocarcinoma/pathology , Female , Gastrointestinal Hormones/metabolism , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Signal Transduction/drug effects , Spheroids, Cellular/drug effects , Spheroids, Cellular/metabolism , Spheroids, Cellular/pathology , Stromal Cells/drug effects , Stromal Cells/metabolism , Trophoblasts/cytology , Vascular Endothelial Growth Factor, Endocrine-Gland-Derived/metabolism , Wound Healing/drug effects , Young Adult
13.
Biol Reprod ; 96(1): 24-33, 2017 01 01.
Article in English | MEDLINE | ID: mdl-28395321

ABSTRACT

Endometrial receptivity is crucial for implantation and establishment of a normal pregnancy. The shift from proliferative to receptive endometrium is still far from being understood. In this paper, we comprehensively present the transcriptome of the human endometrium by comparing endometrial biopsies from proliferative phase with consecutive biopsies 7-9 days after ovulation. The results show a clear difference in expression between the two time points using both total and small RNA sequencing. A total of 3,297 messenger RNAs (mRNAs), 516 long noncoding RNAs (lncRNAs), and 102 small noncoding RNAs were identified as statistically differentially expressed between the two time points. We show a thorough description of the change in mRNA between the two time points and display lncRNAs, small nucleolar RNAs, and small nuclear RNAs not previously reported in the healthy human endometrium. In conclusion, this paper reports in detail the shift in RNA expression from the proliferative to receptive endometrium.


Subject(s)
Endometrium/metabolism , Follicular Phase/metabolism , Luteal Phase/metabolism , RNA/metabolism , Adult , Female , Humans , Primary Cell Culture , Principal Component Analysis , Sequence Analysis, RNA , Young Adult
14.
PLoS One ; 12(1): e0171004, 2017.
Article in English | MEDLINE | ID: mdl-28135285

ABSTRACT

Insulin resistance and compensatory hyperinsulinemia are characteristic features of obesity and polycystic ovary syndrome, and both are associated with reduced fertility and implantation. There is little knowledge about the effect of insulin on the decidualization process and previous findings are contradictory. We investigated the effect of insulin on the regulation of forkhead box protein O1 (FOXO1), one of the most important transcription factors during decidualization. Endometrial stromal cells were isolated from six healthy, regularly menstruating women and decidualized in vitro. Gene expression levels of six putative FOXO1 target genes (including insulin-like growth factor binding protein-1 (IGFBP1) and prolactin (PRL)) were measured with Real-Time PCR following FOXO1 inhibition or insulin treatment. PI3K inhibition was used to identify the possible mechanism behind regulation. Subcellular localization of FOXO1 was analyzed with immunofluorescence. All the genes (IGFBP1, CTGF, INSR, DCN, LEFTY2), except prolactin, were evaluated as FOXO1 target genes in decidualizing stromal cells. Insulin caused a significant dose-dependent inhibition of the verified FOXO1 target genes. It was also demonstrated that insulin regulated FOXO1 target genes by transcriptional inactivation and nuclear export of FOXO1 via PI3K pathway. However, insulin did not inhibit the morphological transformation of endometrial stromal cells via transcriptional inactivation of FOXO1. This study provides new insights on the action of insulin on the endometrium via regulation of FOXO1. It is suggested that hyperinsulinemia results in dysregulation of a high number of FOXO1 controlled genes that may contribute to endometrial dysfunction and reproductive failure. Our findings may illuminate possible reasons for unexplained infertility.


Subject(s)
Decidua/metabolism , Endometrium/cytology , Endometrium/metabolism , Forkhead Box Protein O1/genetics , Insulin/pharmacology , Transcription, Genetic/drug effects , Adolescent , Adult , Bucladesine/pharmacology , Decidua/drug effects , Down-Regulation/drug effects , Down-Regulation/genetics , Endometrium/drug effects , Female , Forkhead Box Protein O1/metabolism , Humans , Medroxyprogesterone Acetate/pharmacology , Stromal Cells/drug effects , Stromal Cells/metabolism , Young Adult
15.
Anticancer Res ; 34(2): 893-8, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24511028

ABSTRACT

BACKGROUND: Kaposi's sarcoma (KS) shows a distinct geographical and ethnic distribution. Genes at both ends of the human herpesvirus 8 (HHV-8) genome have been shown to vary considerably. Seven major molecular subtypes of HHV-8 were defined based on the amino acid sequence of the open reading frame K1 (orf-K1). The aim of the present study was to characterize HHV8 isolates from hospitalized patients in Hungary. MATERIALS AND METHODS: A total of 36 archival paraffin-embedded Kaposi's sarcoma tissue samples were collected. Polymerase chain reaction (PCR) was carried out on the extracted DNA, using specific primers for HHV-8. After identifying the presence of HHV-8 by amplification of its orf26 region, the orf-K1 region was amplified, sequenced and used for phylogenetic analysis. RESULTS: From the 36 orf26-positive cases, orf-K1 was amplified and was analyzed successfully in 12 cases. Phylogenetic studies, based on the complete K1 gene/protein sequences, indicate that all strains belong to the A subtype. Specifically, six of them were related to the A1 subgroup, six to the A2 subgroup and three previously reported to the A3 subgroup. Nucleotide sequence data are reported and are available in the Genbank database under accession numbers KF829938-KF829947.


Subject(s)
Herpesvirus 8, Human/classification , Sarcoma, Kaposi/virology , Aged , Aged, 80 and over , Amino Acid Sequence , Base Sequence , DNA, Viral/blood , DNA, Viral/genetics , Female , Herpesvirus 8, Human/genetics , Herpesvirus 8, Human/isolation & purification , Humans , Hungary/epidemiology , Male , Molecular Epidemiology , Molecular Sequence Data , Molecular Typing , Paraffin Embedding , Phylogeny , Sarcoma, Kaposi/blood , Sarcoma, Kaposi/epidemiology , Sequence Alignment
16.
Proc Natl Acad Sci U S A ; 109(5): 1512-7, 2012 Jan 31.
Article in English | MEDLINE | ID: mdl-22307606

ABSTRACT

Following infection with Epstein-Barr virus (EBV), the virus is carried for life in the memory B-cell compartment in a silent state (latency I/0). These cells do not resemble the proliferating lymphoblastoid cells (LCLs) (latency III) that are generated after infection. It is of fundamental significance to identify how the different EBV expression patterns are established in the latently infected cell. In view of the prompt activatability of CD4(+) T cells in primary EBV infection, and their role in B-cell differentiation, we studied the involvement of CD4(+) T cells in the regulation of EBV latency. Lymphoblastoid cell lines (LCLs) were cocultured with autologous or allogeneic CD4(+) T cells. Activated T cells influenced the expression of two key viral proteins that determine the fate of the infected B cell. EBNA2 was down-regulated, whereas LMP1 was unregulated and the cells proliferated less. This was paralleled by the down-regulation of the latency III promoter (Cp). Experiments performed in the transwell system showed that this change does not require cell contact, but it is mediated by soluble factors. Neutralizing experiments proved that the up-regulation of LMP1 is, to some extent, mediated by IL21, but this cytokine was not responsible for EBNA2 down-regulation. This effect was partly mediated by soluble CD40L. We detected similar regulatory functions of T cells in in vitro-infected lymphocyte populations. In conclusion, our results revealed an additional mechanism by which CD4(+) T cells can control the EBV-induced B-cell proliferation.


Subject(s)
CD4-Positive T-Lymphocytes/metabolism , Herpesvirus 4, Human/physiology , Virus Latency , CD4-Positive T-Lymphocytes/cytology , Coculture Techniques , Humans , Lymphocyte Activation , Solubility
17.
J Virol ; 86(8): 4701-7, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22345482

ABSTRACT

We report that type I interferons (IFNs) upregulate latent membrane protein 1 (LMP-1) expression by direct activation of the ED-L1 promoter in several Epstein-Barr virus (EBV)-carrying Burkitt's lymphoma lines. In EBV-infected primary B cells, IFN-α transiently upregulates LMP-1 mRNA, but not protein levels, followed by downregulation of both, suggesting a novel antiproliferative mechanism of type I IFNs. Furthermore, our results may explain the expression of LMP-1 in memory B cells of systemic lupus erythematosus patients.


Subject(s)
B-Lymphocytes/metabolism , Herpesvirus 4, Human/genetics , Interferon Type I/metabolism , Viral Matrix Proteins/genetics , B-Lymphocytes/drug effects , B-Lymphocytes/virology , Cell Line , Gene Expression Regulation, Viral/drug effects , Herpesvirus 4, Human/immunology , Humans , Interferon Type I/pharmacology , Molecular Sequence Data , Promoter Regions, Genetic , Signal Transduction/drug effects , Transcription, Genetic , Viral Matrix Proteins/metabolism , Virus Latency
18.
Cytokine ; 57(3): 360-71, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22204827

ABSTRACT

Type I interferons (IFN) exert multiple effects on both the innate and adaptive immune system in addition to their antiviral and antiproliferative activities. Little is known, however about the direct effects of type I IFNs on germinal center (GC) B cells, the central components of adaptive B cell responses. We used Burkitt's lymphoma (BL) lines, as a model system of normal human GC B cells, to examine the effect of type I IFNs on the expression of BCL-6, the major regulator of the GC reaction. We show that type I IFNs, but not IFNγ, IL-2 and TNFα rapidly down-regulate BCL-6 protein and mRNA expression, in cell lines derived from endemic, but not from sporadic BL. IFNα-induced down-regulation is specific for BCL-6, independent of Epstein-Barr virus and is not accompanied by IRF-4 up-regulation. IFNα-induced BCL-6 mRNA down-regulation does not require de novo protein synthesis and is specifically inhibited by piceatannol. The proteasome inhibitor MG132 non-specifically prevents, while inhibitors of alternate type I IFN signaling pathways do not inhibit IFNα-induced BCL-6 protein downregulation. We validate our results with showing that IFNα rapidly down-regulates BCL-6 mRNA in purified mouse normal GC B cells. Our results identify type I IFNs as the first group of cytokines that can down-regulate BCL-6 expression directly in GC B cells.


Subject(s)
B-Lymphocytes/metabolism , Burkitt Lymphoma/pathology , DNA-Binding Proteins/metabolism , Down-Regulation/drug effects , Germinal Center/cytology , Interferon-alpha/pharmacology , Adaptor Proteins, Signal Transducing , Animals , B-Lymphocytes/drug effects , Burkitt Lymphoma/immunology , Burkitt Lymphoma/virology , Cell Line, Transformed , DNA-Binding Proteins/genetics , Gene Expression Regulation, Neoplastic/drug effects , Herpesvirus 4, Human/drug effects , Herpesvirus 4, Human/immunology , Humans , Interferon Regulatory Factors/metabolism , Interferon-gamma/pharmacology , Interleukin-2/pharmacology , Kinetics , Leupeptins/pharmacology , Mice , Protein Biosynthesis/drug effects , Proto-Oncogene Proteins c-bcl-6 , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA-Binding Proteins , Signal Transduction/drug effects , Stilbenes/pharmacology , Transcription Factors/metabolism , Tumor Necrosis Factor-alpha/pharmacology
19.
Virus Res ; 120(1-2): 36-48, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16766077

ABSTRACT

The goal of the study was to establish if there was a relationship between molecular patterns and virus evolution. Therefore the complete genome sequence of two distinct apathogenic Newcastle disease virus (NDV) strains was determined and a third genome size category, containing 15,198 nucleotides, was recognized. Phylogenetic analysis revealed that two major separations resulting in three genome size categories occurred during the history of NDV. An ancient division in the primordial reservoir (wild waterbird species) led to two basal sister clades, class I and II, with genome sizes 15,198 (due to a 12 nucleotide insert in the phosphoprotein gene) and 15,186 nucleotides, respectively. Ancestors of only class II viruses colonized chicken populations and subsequently converted to virulent forms. These took place more than once and resulted in an early lineage [including genotypes I-IV and H33(W)] with genome size of 15,186 nucleotides. A second division occurred in the 20th century in the secondary (chicken) host. This gave rise to the branching-off of a clade (including recent genotypes V-VIII consisting of only pathogenic viruses) with the concomitant insertion of six nucleotides into the 5' non-coding region of the nucleoprotein gene thereby increasing the genome size to 15,192 nucleotides.


Subject(s)
Biological Evolution , Genome, Viral , Newcastle Disease/virology , Newcastle disease virus/genetics , Animals , Birds , Molecular Sequence Data , Newcastle disease virus/classification , Newcastle disease virus/pathogenicity , Species Specificity , Virulence
20.
Virus Genes ; 32(1): 49-57, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16525735

ABSTRACT

This paper describes the complete genome sequence of IT-227/82, a strain of avian paramyxovirus type-1 of pigeon (PPMV-1). IT-227/82 is an antigenic variant of Newcastle disease virus (NDV) of chickens. The genome is 15,192 nucleotides (nt) long, similarly to the previously published NDV strain ZJ1. It is, however, six nt longer than the genomes of NDV strains LaSota/46 and Beaudette C. The six-nt insertion was located in the 5' non-coding region of the nucleoprotein (NP) gene. The presence of this six-nt insertion showed no correlation with the virulence or the reservoir of the NDV strains sequenced so far. The genome length of 15,186 or 15,192 nt can be connected however, to "old" (I, II, III and IV) and "new" (V, VI, VII and VIII) NDV genotypes, respectively. Comparison of open reading frames indicated that the PPMV-1 strain IT-227/82 encodes the longest W protein, (227 amino acids). The length of the W protein showed remarkable differences even within genotypes and cannot be regarded, therefore, as a phylogenetic feature. The haemagglutinin-neuraminidase protein of strain IT-227/82 consists of 571 amino acids, similarly to genotypes IV, V and VII NDV strains, while genotype I and II strains have longer HN proteins, 616 and 577 amino acids, respectively. The length of the haemagglutinin-neuraminidase protein possesses phylogenetic importance.


Subject(s)
Columbidae/virology , Newcastle disease virus/genetics , Animals , Base Sequence , Cloning, Molecular , Conserved Sequence , DNA, Viral/genetics , Genome, Viral , Genotype , Molecular Sequence Data , Newcastle disease virus/classification , Newcastle disease virus/immunology , Open Reading Frames , Phylogeny , RNA, Viral/genetics , Species Specificity , Viral Proteins/genetics
SELECTION OF CITATIONS
SEARCH DETAIL
...