Identification of miRNAs and associated pathways regulated by Leukemia Inhibitory Factor in trophoblastic cell lines

ABSTRACTIntroductionLeukemia Inhibitory Factor (LIF) regulates behavior of trophoblast cells and their interaction with immune and endothelial cells.In vitro, trophoblast cell response to LIF may vary depending on the cell model. Reported differences in the miRNA profile of trophoblastic cells may be responsible for these observations. Therefore, miRNA expression was investigated in four trophoblastic cell lines under LIF stimulation followed byin silicoanalysis of altered miRNAs and their associated pathways.MethodsLow density TaqMan miRNA assays were used to quantify levels of 762 mature miRNAs under LIF stimulation in three choriocarcinoma-derived (JEG-3, ACH-3P and AC1-M59) and a trophoblast immortalized (HTR-8/SVneo) cell lines. Expression of selected miRNAs was confirmed in primary trophoblast cells and cell lines by qPCR. Targets and associated pathways of the differentially expressed miRNAs were inferred from the miRTarBase followed by a KEGG Pathway Enrichment Analysis. HTR-8/SVneo and JEG-3 cells were transfected with miR-21-mimics and expression of miR-21 targets was assessed by qPCR.ResultsA similar number of miRNAs changed in each tested cell line upon LIF stimulation, however, low coincidence of individual miRNA species was observed and occurred more often among choriocarcinoma-derived cells (complete data set athttp://www.ncbi.nlm.nih.gov/geo/under GEO accession number GSE130489). Altered miRNAs were categorized into pathways involved in human diseases, cellular processes and signal transduction. Six cascades were identified as significantly enriched, including JAK/STAT and TGFB-SMAD. Upregulation of miR-21-3p was validated in all cell lines and primary cells and STAT3 was confirmed as its target.DiscussionDissimilar miRNA responses may be involved in differences of LIF effects on trophoblastic cell lines.

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Comparison of leukemia inhibitory factor-induced intracellular signalling in different trophoblastic cell lines

Journal of Reproductive Immunology ◽

10.1016/j.jri.2011.06.064 ◽

2011 ◽

Vol 90 (2) ◽

pp. 166

Author(s):

W. Chaiwangyen ◽

F.L. Pereira de Sousa ◽

D.M. Morales Prieto ◽

S. Ospina ◽

U.R. Markert

Keyword(s):

Cell Lines ◽

Leukemia Inhibitory Factor ◽

Intracellular Signalling ◽

Inhibitory Factor ◽

Trophoblastic Cell

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An In Vitro and In Vivo Evidence That Downregulation of Leukemia Inhibitory Factor (LIF) Receptor (LIF-R) Decreases the Metastatic Potential of Human Rhabdomyosarcoma (RMS) Cells.

Blood ◽

10.1182/blood.v108.11.2561.2561 ◽

2006 ◽

Vol 108 (11) ◽

pp. 2561-2561

Author(s):

Marcin Wysoczynski ◽

Katarzyna Miekus ◽

Anna Marcinkowska ◽

Anna Janowska-Wieczorek ◽

Mariusz Z. Ratajczak

Keyword(s):

Skeletal Muscle ◽

Bone Marrow ◽

Cell Line ◽

Cell Lines ◽

Leukemia Inhibitory Factor ◽

Biological Significance ◽

Human Cells ◽

Inhibitory Factor

Abstract Rhabdomyosarcoma (RMS) and skeletal muscle-derived tumors frequently infiltrate bone marrow (BM). We have demonstrated that the stromal-derived factor (SDF)-1-CXCR4 receptor (Blood2002;100:2597) and hepatocyte growth factor (HGF)-c-Met receptor (Cancer Res. 2003;63:7926) play an important role in RMS metastasis to BM. Leukemia inhibitory factor (LIF) is a well known factor that plays an important role in skeletal muscle development/regeneration and similarly as SDF-1 and HGF is secreted by BM stroma. This prompted us to examine whether the LIF-LIF receptor (LIF-R) axis affects the biology/metastasis of RMS cells. We employed in our studies, human established RMS cell lines, as well as RMS samples isolated from patients and noticed that LIF-R was expressed not only on established human RMS cell lines (7/7) but more importantly, it was also detectable in patient samples (23/23). We also found that in RMS cells LIF stimulatesphosphorylation of MAPKp42/44, AKT and STAT3,chemotaxis and adhesion andincreases resistance to cytostatics (e.g., etoposide). These LIF-mediated effects were inhibited after downregulating the LIF-R by siRNA. To learn more on the biological significance of the LIF-LIF-R axis in vivo we employed two models. First, human RMS cells (RH-30) were exposed or not exposed to LIF-R siRNA and subsequently injected into SCID™-Beige immunodeficient mice. To estimate the number of RMS cells that seed to BM and liver in these animals, we isolated DNA and using real- time RT-PCR, amplified human a-satellite sequences and murine b-actin. The number of human cells present in murine organs was subsequently calculated from a standard curve derived from mixing varying numbers of human cells with a constant number of murine cells. We noticed that downregulation of LIF-R by siRNA significantly decreased the number of human RMS cells in murine BM and liver (x4 and x2 respectively). In a second model, the RH30 cell line was selected by repetitive chemotaxis for cells that are highly responsive to LIF (RH-30 L) and subsequently the cells from parental RH-30 cell line and RH-30 L cells were injected intramuscularly. Six weeks after tumour inoculation, we detected more metastasis in bone marrow and lungs in mice injected with RH-30L cells as compared to parental RH-30 clone (x6 and x3 respectively). In conclusion, we present evidence for the first time that the inhibition of LIF-LIF-R axis may decrease the invasive potential of human RMS both in vitro and in vivo. Hence, molecular targeting of LIF-LIF-R axis could possibly become a more effective new strategy to control the progression and metastasis of RMS.

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Identification of miRNAs and associated pathways regulated by Leukemia Inhibitory Factor in trophoblastic cell lines

Placenta ◽

10.1016/j.placenta.2019.09.005 ◽

2019 ◽

Vol 88 ◽

pp. 20-27 ◽

Cited By ~ 3

Author(s):

Diana M. Morales-Prieto ◽

Emanuel Barth ◽

Jose Martín Murrieta-Coxca ◽

Rodolfo R. Favaro ◽

Ruby N. Gutiérrez-Samudio ◽

...

Keyword(s):

Cell Lines ◽

Leukemia Inhibitory Factor ◽

Inhibitory Factor ◽

Trophoblastic Cell

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Transcriptomic analysis of primate placentas and novel rhesus trophoblast cell lines informs investigations of human placentation

10.1101/2020.08.21.262030 ◽

2020 ◽

Author(s):

Jimi L. Rosenkrantz ◽

Jessica E. Gaffney ◽

Victoria HJ. Roberts ◽

Lucia Carbone ◽

Shawn L. Chavez

Keyword(s):

Animal Model ◽

Cell Lines ◽

Pregnancy Complications ◽

Rhesus Macaques ◽

Enrichment Analysis ◽

Well Being ◽

First Trimester ◽

Trophoblast Cell ◽

Pathway Enrichment Analysis

AbstractProper placentation, including trophoblast differentiation and function, is essential for the health and well-being of both the mother and baby throughout pregnancy. Placental abnormalities that occur during the early stages of development are thought to contribute to pre-eclampsia and other placenta-related pregnancy complications. However, relatively little is known about these stages in humans due to obvious ethical and technical limitations. Rhesus macaques are considered an ideal surrogate for studying human placentation, but the unclear translatability of known human placental markers and lack of accessible rhesus trophoblast cell lines can impede the use of this animal model. Here, we performed a cross-species transcriptomic comparison of human and rhesus placenta and determined that while the majority of known placental markers were similarly expressed, 952 differentially expressed genes (DEGs) were identified between the two species. Pathway enrichment analysis of the 447 human-upregulated DEGs, including ADAM12, ERVW-1, KISS1, LGALS13, PAPPA2, PGF, and SIGLEC6, revealed over-representation of functional terms associated with pre-eclampsia and other pregnancy disorders. Additionally, to enable in vitro functional studies of early placentation, we generated and thoroughly characterized two highly-pure first-trimester telomerase (TERT) immortalized rhesus trophoblast cell lines (iRP-D26 and iRP-D28A) that retained crucial features of isolated primary trophoblasts. Overall, our findings help elucidate the molecular translatability between human and rhesus placenta and reveal notable expression differences in human placental markers and genes associated with pregnancy complications that should be considered when using the rhesus animal model to study normal and pathological human placentation.

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YB-1 Is Altered in Pregnancy-Associated Disorders and Affects Trophoblast in Vitro Properties via Alternation of Multiple Molecular Traits

International Journal of Molecular Sciences ◽

10.3390/ijms22137226 ◽

2021 ◽

Vol 22 (13) ◽

pp. 7226

Author(s):

Violeta Stojanovska ◽

Aneri Shah ◽

Katja Woidacki ◽

Florence Fischer ◽

Mario Bauer ◽

...

Keyword(s):

Cell Lines ◽

Cancer Progression ◽

Cell Function ◽

Trophoblast Cell ◽

Mrna Levels ◽

Trophoblast Cells ◽

Invasion And Migration ◽

And Migration ◽

Apoptosis And Inflammation

Cold shock Y-box binding protein-1 (YB-1) coordinates several molecular processes between the nucleus and the cytoplasm and plays a crucial role in cell function. Moreover, it is involved in cancer progression, invasion, and metastasis. As trophoblast cells share similar characteristics with cancer cells, we hypothesized that YB-1 might also be necessary for trophoblast functionality. In samples of patients with intrauterine growth restriction, YB-1 mRNA levels were decreased, while they were increased in preeclampsia and unchanged in spontaneous abortions when compared to normal pregnant controls. Studies with overexpression and downregulation of YB-1 were performed to assess the key trophoblast processes in two trophoblast cell lines HTR8/SVneo and JEG3. Overexpression of YB-1 or exposure of trophoblast cells to recombinant YB-1 caused enhanced proliferation, while knockdown of YB-1 lead to proliferative disadvantage in JEG3 or HTR8/SVneo cells. The invasion and migration properties were affected at different degrees among the trophoblast cell lines. Trophoblast expression of genes mediating migration, invasion, apoptosis, and inflammation was altered upon YB-1 downregulation. Moreover, IL-6 secretion was excessively increased in HTR8/SVneo. Ultimately, YB-1 directly binds to NF-κB enhancer mark in HTR8/SVneo cells. Our data show that YB-1 protein is important for trophoblast cell functioning and, when downregulated, leads to trophoblast disadvantage that at least in part is mediated by NF-κB.

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Lipidomics profiling of goose granulosa cell model of stearoyl-CoA desaturase function identifies a pattern of lipid droplets associated with follicle development

Cell & Bioscience ◽

10.1186/s13578-021-00604-6 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Xin Yuan ◽

Shenqiang Hu ◽

Liang Li ◽

Chunchun Han ◽

Hehe Liu ◽

...

Keyword(s):

Differentially Expressed Genes ◽

Lipid Droplets ◽

Cell Model ◽

Enrichment Analysis ◽

Differentially Expressed ◽

Pathway Enrichment Analysis ◽

Follicle Development ◽

Microscopy Analysis ◽

Stearoyl Coa Desaturase ◽

Underlying Mechanisms

Abstract Background Despite their important functions and nearly ubiquitous presence in cells, an understanding of the biology of intracellular lipid droplets (LDs) in goose follicle development remains limited. An integrated study of lipidomic and transcriptomic analyses was performed in a cellular model of stearoyl-CoA desaturase (SCD) function, to determine the effects of intracellular LDs on follicle development in geese. Results Numerous internalized LDs, which were generally spherical in shape, were dispersed throughout the cytoplasm of granulosa cells (GCs), as determined using confocal microscopy analysis, with altered SCD expression affecting LD content. GC lipidomic profiling showed that the majority of the differentially abundant lipid classes were glycerophospholipids, including PA, PC, PE, PG, PI, and PS, and glycerolipids, including DG and TG, which enriched glycerophospholipid, sphingolipid, and glycerolipid metabolisms. Furthermore, transcriptomics identified differentially expressed genes (DEGs), some of which were assigned to lipid-related Gene Ontology slim terms. More DEGs were assigned in the SCD-knockdown group than in the SCD-overexpression group. Integration of the significant differentially expressed genes and lipids based on pathway enrichment analysis identified potentially targetable pathways related to glycerolipid/glycerophospholipid metabolism. Conclusions This study demonstrated the importance of lipids in understanding follicle development, thus providing a potential foundation to decipher the underlying mechanisms of lipid-mediated follicle development.

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Endometrial leukemia inhibitory factor as a predictor of pregnancy after in vitro fertilization

International Journal of Gynecology & Obstetrics ◽

10.1016/j.ijgo.2007.12.005 ◽

2008 ◽

Vol 102 (1) ◽

pp. 23-27 ◽

Cited By ~ 10

Author(s):

Paulo Serafini ◽

André M. Rocha ◽

Cyntia T. Osório ◽

Ismael da Silva ◽

Eduardo L. Motta ◽

...

Keyword(s):

In Vitro Fertilization ◽

Leukemia Inhibitory Factor ◽

Inhibitory Factor ◽

Vitro Fertilization

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Comparative pathway enrichment analysis in gastrointestinal cell lines Caco-2, HT-29, HEPG2, and colon fibroblasts using a custom expression panel for tight-junction and cytoskeletal regulatory genes

10.1101/747105 ◽

2019 ◽

Cited By ~ 1

Author(s):

JM Robinson

Keyword(s):

Gene Expression ◽

Comparative Analysis ◽

Tight Junction ◽

Cell Lines ◽

Enrichment Analysis ◽

Pathway Enrichment Analysis ◽

Intestinal Epithelial ◽

Expression Data ◽

Pathway Enrichment ◽

Ht 29

AbstractThis brief report details results from a comparative analysis of Nanostring expression data between cell lines HEPG2, Caco-2, HT-29, and colon fibroblasts. Raw and normalized data are available publicly in the NCBI GEO/Bioproject databases. Results identify cell-line specific variations in gene expression relevant to intestinal epithelial function.

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High expression of MCM10 is predictive of poor outcomes in lung adenocarcinoma

PeerJ ◽

10.7717/peerj.10560 ◽

2021 ◽

Vol 9 ◽

pp. e10560

Author(s):

Mingrui Shao ◽

Shize Yang ◽

Siyuan Dong

Keyword(s):

Lung Adenocarcinoma ◽

Complex Disease ◽

Enrichment Analysis ◽

Integrated Approach ◽

Replication Initiation ◽

Prognostic Biomarkers ◽

Normal Lung ◽

Pathway Enrichment Analysis ◽

Therapeutic Value

Backgrounds Lung adenocarcinoma is a complex disease that results in over 1.8 million deaths a year. Recent advancements in treating and managing lung adenocarcinoma have led to modest decreases in associated mortality rates, owing in part to the multifactorial etiology of the disease. Novel prognostic biomarkers are needed to accurately stage the disease and act as the basis of adjuvant treatments. Material and Methods The microarray datasets GSE75037, GSE31210 and GSE32863 were downloaded from the Gene Expression Omnibus (GEO) database to identify prognostic biomarkers for lung adenocarcinoma and therapy. The differentially expressed genes (DEGs) were identified by GEO2R. Functional and pathway enrichment analysis were performed by Kyoto Encyclopedia of Genes and Genomes and Gene Ontology (GO). Validation was performed based on 72 pairs of lung adenocarcinoma and adjacent normal lung tissues. Results Results showed that the DEGs were mainly focused on cell cycle and DNA replication initiation. Forty-one hub genes were identified and further analyzed by CytoScape. Here, we provide evidence which suggests MCM10 is a potential target with prognostic, diagnostic and therapeutic value. We base this on an integrated approach of comprehensive bioinformatics analysis and in vitro validation using the A549 lung adenocarcinoma cell line. We show that MCM10 overexpression correlates with a poor prognosis, while silencing of this gene decreases aberrant growth by 2-fold. Finally, evaluation of 72 clinical biopsy samples suggests that overexpression of MCM10 in the lung adenocarcinoma highly correlates with larger tumor size. Together, this work suggests that MCM10 may be a clinically relevant gene with both predictive and therapeutic value in lung adenocarcinoma.

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