miR-3074-5p/CLN8 pathway regulates decidualization in recurrent miscarriage

Reproduction ◽  
2021 ◽  
Vol 162 (1) ◽  
pp. 33-45
Author(s):  
Nan Meng ◽  
Xinyue Wang ◽  
Yan Shi ◽  
Yanyan Mao ◽  
Qian Yang ◽  
...  
Keyword(s):  
Early Pregnancy ◽  
Recurrent Miscarriage ◽  
Cyclin B1 ◽  
Expression Level ◽  
Extravillous Trophoblast ◽  
Control Group ◽  
Endometrial Stromal Cells ◽  
Early Pregnancy Failure ◽  
Intracellular Expression

Decidualization is essential for the successful establishment of pregnancy, and the dysregulated decidualization may lead to early pregnancy loss. It was previously reported by us that miR-3074-5p could promote apoptosis but inhibit invasion of human extravillous trophoblast (EVT) cells in vitro, and the expression level of miR-3074-5p in villus tissues of recurrent miscarriage (RM) patients was significantly increased. The aim of this study was to preliminarily explore the role of miR-3074-5p played in the decidualization of human endometrial stromal cells (ESCs). It was found that the decidual expression level of miR-3074-5p in RM patients was remarkably higher than that in the control group. The overexpression of miR-3074-5p in the immortalized human ESC line, T-HESCs, showed suppressive effects not only on the cell proliferation, as well as the intracellular expression levels of cyclin B1 (CCNB1), CCND1 and CCNE1 but also on the in vitro-induced decidualization. CLN8 mRNA, encoding an endoplasmic reticulum (ER)-associated membrane protein, was validated to be directly targeted by miR-3074-5p. And, the expression level of CLN8 was continuously increased along with the decidualization process, whereas down-regulated CLN8 expression could inhibit the decidualization of T-HESCs in vitro. Furthermore, contrary to the increased expression level of miR-3074-5p, a significantly decreased CLN8 expression was observed in decidual tissues of RM patients. Collectively, these data suggested that an increased miR-3074-5p expression in ESCs might cause early pregnancy failure by disturbing decidualization of ESCs via the miR-3074-5p/CLN8 pathway, providing a potential diagnostic and therapeutic target for RM.

MNSFβ Promotes the Proliferation and Migration of Human Extravillous Trophoblast Cells and the Villus Expression Level of MNSFβ Is Decreased in Recurrent Miscarriage Patients

2020 ◽  
Vol 86 (1-2) ◽  
pp. 27-39
Author(s):  
Nan Wang ◽  
Qian Yang ◽  
Yan Gu ◽  
Xingxing Zhen ◽  
Yan Shi ◽  
...  
Keyword(s):  
Early Pregnancy ◽  
Recurrent Miscarriage ◽  
First Trimester ◽  
Expression Level ◽  
Extravillous Trophoblast ◽  
Placental Villus ◽  
Expression Levels ◽  
Placental Villi ◽  
And Migration ◽  
Proliferation And Migration

<b><i>Aims:</i></b> The invasion of extravillous trophoblast (EVT) cells into maternal decidua is essential for the establishment and maintenance of pregnancy. Derangement of EVT cell invasion might cause pregnancy complications including recurrent miscarriage (RM). We previously reported that deficiency of monoclonal nonspecific suppressor factor beta (MNSFβ) led to the early pregnancy failure in mice and the decidual MNSFβ expression level in RM patients was significantly decreased, but the underlying molecular mechanism of the role that MNSFβ played at the maternal-fetal interface remains unclear. Thus, in the present study, we determined effects of downregulated MNSFβ expression on human EVT cell activities. <b><i>Methods:</i></b> The MNSFβ expression in first-trimester human decidual and placental villus tissues was detected, respectively, by immunofluorescence or immunohistochemical analyses. The MNSFβ expression level in the immortalized first-trimester human EVT cell line HTR8/SVneo was downregulated by transfecting the small interfering RNA against MNSFβ and upregulated by transfecting the recombinant pDsRed-MNSFβ plasmids. The proliferation, migration, invasion, and apoptosis activities of HTR8/SVneo cells were, respectively, determined by cytometry assay, scratch test, transwell assay, and FITC/PI staining. The expression levels of P53, RhoA, Bcl-2, Bax, and MMP-9 in HTR8/SVneo cells, as well as the expression levels of MNSFβ and RhoA in placental villi of RM patients and physically normal pregnant women (NP), were examined by Western blot analysis. <b><i>Results:</i></b> MNSFβ protein signals were observed in first-trimester human villus and extravillous trophoblast cells. The downregulated MNSFβ expression significantly attenuated the proliferation, migration, and invasion abilities of HTR8/SVneo cells, accompanied with the obviously decreased expression levels of P53, RhoA, Bcl-2, Bax, and MMP-9, whereas the upregulated MNSFβ expression in HTR8/SVneo cells represented the inverse effects. Furthermore, expression levels of MNSFβ and RhoA in first-trimester human placental villus tissues of RM patients were significantly decreased compared to that of NP women. <b><i>Conclusion:</i></b> These data suggested that MNSFβ promotes proliferation and migration of human EVT cells, probably via the P53 signaling pathway, and the deficiency of MNSFβ in placental villi might lead to early pregnancy loss by reducing proliferation and invasion activities of EVTs.

scholarly journals Ghrelin Is Involved in the Decidualization of Human Endometrial Stromal Cells

2003 ◽  
Vol 88 (5) ◽  
pp. 2335-2340 ◽  
Author(s):  
Keisuke Tanaka ◽  
Hiroyuki Minoura ◽  
Tetsuya Isobe ◽  
Hitoshi Yonaha ◽  
Hiroaki Kawato ◽  
...  
Keyword(s):  
Menstrual Cycle ◽  
Early Pregnancy ◽  
Stromal Cells ◽  
Immunohistochemical Analysis ◽  
First Trimester ◽  
Extravillous Trophoblast ◽  
Endometrial Stromal Cells ◽  
Decidual Cells ◽  
Normal Menstrual Cycle

Successful implantation involves a complex interaction between the endometrium and the embryo. It is well known that several neuropeptides are expressed in the endometrium and placenta during embryonal implantation, suggesting an important role as chemical mediators of the feto-maternal relationship. Ghrelin has recently been identified as the endogenous ligand for the GH secretagogue receptor. Ghrelin is a peptide hormone with many physiological functions, and its expression in the human placenta has been reported. To investigate the involvement of ghrelin in embryonal implantation, we assessed the spatio-temporal expression pattern of ghrelin and its receptor in the human endometrium and placenta through the normal menstrual cycle and in early pregnancy. We also examined the effect of ghrelin on the decidualization of endometrial stromal cells (ESC). Weak expression of ghrelin mRNA was detected in the nonpregnant endometrium, and it was dramatically increased in the decidualized endometrium. A GH secretagogue receptor mRNA was detected in the endometrium throughout the normal menstrual cycle and in early pregnancy, but not in the first trimester placenta. Immunohistochemical analysis using an antighrelin antibody revealed strong signals in decidual cells and extravillous trophoblast cells. Coculture with first trimester placenta up-regulated ghrelin mRNA expression by primary cultured ESC, although sex steroids and 8-bromo-cAMP had no effect. In addition, ghrelin enhanced the decidualization of ESC induced by 8-bromo-cAMP (8-Br-cAMP) in vitro. Thus, ghrelin is a novel paracrine/autocrine factor that is involved in cross-talk between the endometrium and embryo during embryonal implantation.

Levels interleukine-4 and interleukin-17 (IL-4, IL-17) of blood in patients with habitual recurrent pregnancy loss, which occurred in a loop in vitro fertilization

2016 ◽  
pp. 137-139
Author(s):  
K.P. Golovatyuk ◽  
Keyword(s):  
In Vitro Fertilization ◽  
Conditioned Medium ◽  
Mononuclear Cells ◽  
Recurrent Miscarriage ◽  
Interleukin 4 ◽  
Control Group ◽  
Interleukin 17 ◽  
Vitro Fertilization ◽  
Blood Mononuclear Cells

The objective: was to investigate the levels of cytokines IL-4 and IL-17 in serum and conditioned medium cultures of blood mononuclear cells (MNC) and evaluation association between their products and miscarriage, which occurred in IVF cycles. Patients and methods. We observed 240 patients with recurrent miscarriage, came in IVF cycles, and 100 apparently healthy fertile women in the control group. The concentrations of IL-4 and IL-17 in serum and conditioned medium of MNC cultures were determined. Results. The levels of IL-4 in the serum and conditioned medium in spontaneous and stimulated mitogen secretion was not significantly different from those in the control group, whereas IL-17 levels were higher than those in the control group serum, in conditioned media of stimulated and non-stimulated MNCs. Conclusion. Disregulation of activity of circulating blood mononuclear cells in women with recurrent miscarriage that followed IVF, is accompanied by increased secretion of IL-17 and almost constant production of IL-4 on the back of high stimulation index of production of these cytokines. Key words: in vitro fertilization, miscarriage, interleukin-4, interleukin-17, serum stimulated and non-stimulated mononuclear blood.

scholarly journals MiR-206 regulates the progression of osteoporosis via targeting HDAC4

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Zhiyuan Lu ◽  
Dawei Wang ◽  
Xuming Wang ◽  
Jilong Zou ◽  
Jiabing Sun ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cell Apoptosis ◽  
Diagnostic Value ◽  
Expression Level ◽  
Luciferase Reporter ◽  
Control Group ◽  
Luciferase Reporter Gene ◽  
Mineral Density ◽  
Gene Assay

Abstract Background More and more studies have confirmed that miRNAs play an important role in maintaining bone remodeling and bone metabolism. This study investigated the expression level of miR-206 in the serum of osteoporosis (OP) patients and explored the effect and mechanism of miR-206 on the occurrence and development of osteoporosis. Methods 120 postmenopausal women were recruited, including 63 cases with OP and 57 women without OP. The levels of miR-206 were determined by qRT-PCR technology. Spearman correlation coefficient was used to evaluate the correlation of miR-206 with bone mineral density (BMD). An ROC curve was used to evaluate the diagnostic value of miR-206 in osteoporosis. The effects of miR-206 on cell proliferation and cell apoptosis of hFOBs were measured by CCK-8 assay and flow cytometry, respectively. Luciferase reporter gene assay was used to confirm the interaction of miR-206 and the 3′UTR of HDAC4. Results Serum miR-206 had low expression level in osteoporosis patient group compared with control group. The expression level of serum miR-206 had diagnostic value for osteoporosis, and the serum miR-206 levels were positively correlated with BMD. The down-regulated miR-206 could inhibit cell proliferation and promote cell apoptosis. Luciferase analysis indicated that HDAC4 was the target gene of miR-206. Conclusions MiR-206 could be used as a new potential diagnostic biomarker for osteoporosis, and in in vitro cell experiments, miR-206 may regulate osteoblast cell proliferation and apoptosis by targeting HDAC4.

scholarly journals Menstrual flow as a non-invasive source of endometrial organoids

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Tereza Cindrova-Davies ◽  
Xiaohui Zhao ◽  
Kay Elder ◽  
Carolyn J. P. Jones ◽  
Ashley Moffett ◽  
...  
Keyword(s):  
Early Pregnancy ◽  
Recurrent Miscarriage ◽  
Milk Proteins ◽  
Receptor Expression ◽  
In Vitro Fertilisation ◽  
Non Invasive ◽  
Menstrual Flow ◽  
Pregnancy Hormones

AbstractAssessment of the endometrium often necessitates a biopsy, which currently involves an invasive, transcervical procedure. Here, we present an alternative technique based on deriving organoids from menstrual flow. We demonstrate that organoids can be derived from gland fragments recovered from menstrual flow. To confirm they faithfully reflect the in vivo state we compared organoids derived from paired scratch biopsies and ensuing menstrual flow from patients undergoing in vitro fertilisation (IVF). We demonstrate that the two sets of organoids share the same transcriptome signature, derivation efficiency and proliferation rate. Furthermore, they respond similarly to sex steroids and early-pregnancy hormones, with changes in morphology, receptor expression, and production of ‘uterine milk’ proteins that mimic those during the late-secretory phase and early pregnancy. This technique has wide-ranging impact for non-invasive investigation and personalised approaches to treatment of common gynaecological conditions, such as endometriosis, and reproductive disorders, including failed implantation after IVF and recurrent miscarriage.

Heparin and aspirin attenuate placental apoptosis in vitro: Implications for early pregnancy failure

2005 ◽  
Vol 192 (1) ◽  
pp. 23-30 ◽  
Author(s):  
Patrick Bose ◽  
Simon Black ◽  
Mamed Kadyrov ◽  
Ute Weissenborn ◽  
Joseph Neulen ◽  
...  
Keyword(s):  
Early Pregnancy ◽  
Pregnancy Failure ◽  
Early Pregnancy Failure

scholarly journals Downregulation of decidual SKP2 is associated with human recurrent miscarriage

2021 ◽  
Author(s):  
Shijian Lv ◽  
Mei Liu ◽  
Lizhen Xu ◽  
Cong Zhang
Keyword(s):  
Stromal Cells ◽  
Recurrent Miscarriage ◽  
Aerobic Glycolysis ◽  
Critical Role ◽  
Protein S ◽  
Pcr Analysis ◽  
Endometrial Stromal Cells ◽  
Downstream Target ◽  
F Box Protein

Abstract Background: Recurrent miscarriage (RM) is a very frustrating problem for both couples and clinicians. To date, the etiology of RM remains poorly understood. Decidualization plays a critical role in implantation and the maintenance of pregnancy, and its deficiency is closely correlated with RM. The F-box protein S-phase kinase associated protein 2 (SKP2) is a key component of the SCF-type E3 ubiquitin ligase complex, which is critically involved in ErbB family-induced Akt ubiquitination, aerobic glycolysis and tumorigenesis. SKP2 is pivotal for reproduction, and SKP2-deficient mice show impaired ovarian development and reduced fertility.Methods: Here, we investigated the expression and function of SKP2 in human decidualization and its relation with RM. A total of 40 decidual samples were collected. Quantitative PCR analysis, western blot analysis and immunohistochemistry analysis were performed to analyze the differential expression of SKP2 between RM and control cells. For in vitro induction of decidualization, both HESCs (human endometrial stromal cells) cell line and primary ESCs (endometrial stromal cells) were used to analyze the effects of SKP2 on decidualization via siRNA transfection.Results: Compared to normal pregnant women, the expression of SKP2 was reduced in the decidual tissues from individuals with RM. After in vitro induction of decidualization, knockdown of SKP2 apparently attenuated the decidualization of HESCs and resulted in the downregulation of HOXA10 and FOXM1, which are essential for normal human decidualization. Moreover, our experiments demonstrated that SKP2 silencing reduced the expression of its downstream target GLUT1.Conclusions: Our study indicates a functional role of SKP2 in RM: downregulation of SKP2 in RM leads to impaired decidualization and downregulation of GLUT1 and consequently predisposes individuals to RM.

scholarly journals Increased expression of HMGB1 in the implantation phase endometrium is related to recurrent implantation failure

2021 ◽  
Author(s):  
Mi Han ◽  
Yi Cao ◽  
Wenjie Zhou ◽  
Mingjuan Zhou ◽  
Xiaowei Zhou ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Underlying Mechanism ◽  
Endometrial Receptivity ◽  
In Vitro Assays ◽  
Control Group ◽  
Implantation Failure ◽  
Recurrent Implantation Failure ◽  
Tubal Infertility ◽  
Endometrial Stromal Cells

Abstract Impaired endometrial receptivity is the main cause of recurrent implantation failure (RIF), however, its underlying mechanism is unclear. In this study, we found that HMGB1 expression was significantly decreased in the implantation phase endometrium in the control group (patients with tubal infertility who successfully achieved conception after the first embryo transfer) (P = 0.006). However, the expression levels of HMGB1 mRNA and protein were significantly upregulated during the implantation phase in endometrial tissues obtained from patients with RIF compared to those in the control group (P = 0.001), consistent with the results of genome-wide expression profiling. Moreover, in vitro assays showed that increased expression of HMGB1 in human endometrial epithelial cells cause marked deficiency in supporting blastocysts and human embryonic JAR cell adhesion, mimicking the process of embryo adhesion. However, overexpression of HMGB1 had no effect on cell proliferation and in-vitro decidualization in a human endometrial stromal cell line (T-HESCs) and in primary human endometrial stromal cells (HESCs). These findings indicate that increased HMGB1 levels suppressed the adhesion capability of epithelial cells, contributing to impaired endometrial receptivity in patients with recurrent implantation failure. This characteristic can be used as a target for detecting and treating recurrent implantation failure in clinical practice.

scholarly journals Comparison of ADAM17 gene expression level in acute lymphoblastic leukemia patients

2021 ◽  
Vol 23 (2) ◽  
pp. 76-80
Author(s):  
Shakiba Karimi ◽  
Noosha Zia Jahromi
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Lymphoblastic Leukemia ◽  
Venous Blood ◽  
Expression Level ◽  
Control Group ◽  
Healthy Individuals ◽  
Blood Samples ◽  
Gapdh Gene ◽  
Large Numbers

Background and aims: In acute lymphoblastic leukemia (ALL), large numbers of stem cells become lymphoblasts or lymphocytes. Among the genetic factors influencing cancer is the ADAM (a disintegrin and metalloprotease) gene family. Due to the important role of this family in cancer, this study aimed to compare the expression level of ADAM17 gene in patients with ALL and healthy individuals. Material and Methods: In this case-control study, 40 venous blood samples were taken from ALL patients referred to Omid hospital in Isfahan, Iran. Also, 40 venous blood samples were taken from healthy individuals in vitro. Lymphocyte isolation was performed using a ficol and cell RNA was isolated using an RNX-Plus kit. It was then converted to cDNA using the Yekta Tajhiz Azma kit. Finally, reverse transcription-polymerase chain reaction (RT-PCR) technique was used to evaluate the relative expression of ADAM17 gene in blood samples of healthy individuals and patients with leukemia, and the ratio was measured with the reference GAPDH gene. SPSS software version 22 and t test were used to analyze the data. Results: The expression level of ADAM17 gene in patients with ALL compared to the control group showed a significant increase, which was statistically significant (P=0.043). Conclusion: It seems that increasing the expression of ADAM17 gene in people with ALL is a suitable biomarker to diagnose this disease.

289 THE EFFECTS OF MEIOSIS BLOCKING BY CDK INHIBITORS ON THE ACTIVITY OF MATURATION PROMOTING FACTOR, ERK1 AND 2, AND THE DISTRIBUTION OF CYTOPLASMIC ORGANELLES IN IN VITRO-MATURED BOVINE OOCYTES

2015 ◽  
Vol 27 (1) ◽  
pp. 233
Author(s):  
R. R. D. Maziero ◽  
C. R. F. Guaitolini ◽  
D. M. Paschoal ◽  
A. M. Crespilho ◽  
F. C. Landim-Alvarenga
Keyword(s):  
Cyclin B1 ◽  
Oocyte Quality ◽  
Control Group ◽  
Cdk Inhibitors ◽  
Cortical Granules ◽  
Immature Oocytes ◽  
Nuclear Maturation ◽  
Cytoplasmic Organelles ◽  
Bovine Oocytes

Studies have suggested that the prematuration with meiotic blockers can improve oocyte quality promoting embryonic development; however, its exact effects on cytoplasmic characteristics remain unclear. Thus, this study aimed to evaluate the effects of meiosis block of bovine oocytes with the CDK inhibitors roscovitine (ROS) and butyrolactone (BL-I) on nuclear maturation, expression, and localization of ERK 1 and 2, cyclin B1, and p34cdc2 proteins and the ultrastructure of oocytes. Immature oocytes from the slaughterhouse were divided into the following groups: (1) control, in vitro maturated (IVM) in TCM 199 for 24 h; (2) ROS 12.5 µM; (3) BL-I 50 µM; and (4) ROS (6.25 µM) + BL-I (25 µM) treatment for 6 h followed by IVM in CDK inhibitor-free medium for 18 h. Immature oocytes and IVM oocytes in each group were then fixed stained by immunofluorescence for nuclear visualisation (n = 600), localization, and expression of ERK1 and 2 proteins, cyclin B1 and p34cdc2 protein (n = 350), and prepared for evaluation of the ultrastructure by electron microscopy (n = 100). Data were analysed using the ANOVA test (nuclear visualisation), Student-Newman-Keuls test (expression of ERK1 and 2 proteins, cyclin B1 and p43cdc2) by the PROC GLM procedure of SAS (SAS Inst. Inc., Cary, NC, USA). At 6 h of IVM, a lower (P < 0.05) percentage of oocytes were at the metaphase I (MI) stage in the control group (C = 18.2 ± 5.4%) compared with other groups and a higher percentage of oocytes were degenerated in the ROS group (16.3 ± 5.6%) compared with other groups (C = 13.6 ± 4.6%, BL-I = 10.0 ± 4.5%, BL-I+ROS = 8.0 ± 5.6%). At 24 h of IVM, higher (P < 0.05) percentages of oocytes were at the MI stage in the control and ROS group (8.3 ± 5.9% and 6.8 ± 6.4%, respectively). There was no difference (P > 0.05) in percentage of metaphase II (MII) oocytes among the groups. Only the ROS group showed lower fluorescence intensity of ERK1 and 2 proteins in the ooplasma (P < 0.05). Immature oocytes showed higher expression of cyclin B1 and p34cdc2 (P < 0.05). There was no difference in the localization of these proteins in the ooplasm and there was no difference in the oocyte ultrastructure (mitochondria, cortical granules, endoplasmic reticulum, microvilli, zona pellucida, lipid granules) among treatments (P > 0.05). The results suggest that a temporary blocking of oocyte maturation by CDK inhibitors affect neither the expression and distribution of MPF components (cyclin B1/cdc2) nor the distribution of cytoplasmic organelles in IVM oocytes. However, the expression of ERK 1 and 2 in mature oocytes may be reduced by pre-IVM with ROS.

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