Differential expression of microRNAs in the placentae of Chinese patients with severe pre-eclampsia
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ABSTRACT: To determine differential expression of microRNAs in placentae with severe pre-eclampsia and normal placenta. Differential expression of microRNAs in placentae (4 severe pre-eclampsia and 4 normal control) was screened by microarray platform, then some differential microRNAs were selected and validated with real-time quantitative reverse transcription polymerase chain reaction in placentae of severe pre-eclampsia (n=24) and normal control group (n=26). Results: We validated the expression of some microRNAs altered in the microarray, and found the following microRNAs were significantly increased in severe pre-eclampsia placentae: miR-16, miR-29b, miR-195, miR-26b, miR-181a, miR-335 and miR-222. Conclusion: These differential microRNAs may play an important role in pathogenesis of pre-eclampsia. We analyzed the microRNA expression in the placentae of Chinese patients with severe pre-eclampsia.
Project description:To determine differential expression of microRNAs in placentae with severe pre-eclampsia and normal placenta. Differential expression of microRNAs in placentae (4 severe pre-eclampsia and 4 normal control) was screened by microarray platform, then some differential microRNAs were selected and validated with real-time quantitative reverse transcription polymerase chain reaction in placentae of severe pre-eclampsia (n=24) and normal control group (n=26). Results: We validated the expression of some microRNAs altered in the microarray, and found the following microRNAs were significantly increased in severe pre-eclampsia placentae: miR-16, miR-29b, miR-195, miR-26b, miR-181a, miR-335 and miR-222. Conclusion: These differential microRNAs may play an important role in pathogenesis of pre-eclampsia.
Project description:Latexin (Lxn) was originally isolated as a tissue specific marker of rat neuron. It is expressed in various tissues of humans and many other vertebrates. Lxn inhibits human carboxypeptidase A4 (hCPA4), whose expression is induced by histone deacetylase inhibitors in prostate cancer cells, and is associated with cancer progression. Structural analysis has shown significant similarity between Lxn protein and the tumor suppressor protein TIG1. Recent reports have demonstrated Lxn functions in the negative control of hematopoietic stem cell numbers (HSC) in mice. In the present study, an anti-Lxn monoclonal antibody 1G11 was generated and Western blot analysis showed that Lxn was expressed mainly in the cytoplasm of numerous human cell lines. Immunohistochemical analysis in gastric cancer tissues and the corresponding adjacent normal tissues showed that Lxn expression was lower in cancer tissues as compared to normal tissues. In addition, the Lxn gene was exogenously introduced into a Lxn null gastric cancer cell line, MGC803. Results of colony formation assay, soft agar assay and tumor growth in nude mice showed that over-expression of Lxn inhibits the proliferation and tumorigenicity of MGC803 cells expressing Lxn. Thirty-seven genes, whose expression were altered in response to Lxn expression were identified by microarray analysis. Methylation analysis of the central promoter region of the Lxn gene in several cell lines suggested that Lxn expression was silenced by CpG hypermethylation. Taken together, our results indicate that Lxn is a potential tumor suppressor and plays a role in negative control of tumor cell growth. We analyzed gene expression differece between Latexin (Lxn) postive cell line and negtive cells to find the active pathway and related biology function.
Project description:Hypertensive disorders in pregnancy, of which the multisystem pathology pre-eclampsia is most severe, often lead to preterm delivery, maternal mortality and life-long complications. Pre-eclampsia lacks early screening tools and causal therapies, illustrating the urgent need for a better understanding of early disease dynamics. Here, we present the first study comparing single-nuclei transcriptomes of human diseased preterm preeclamptic placentae and healthy controls, embedding the characterization of the maternal-fetal barrier dysfunction in the context of a comprehensive spatio-temporal study including early and late gestational placentae. Our results highlight and contextualize a perturbed communication from fetal to maternal side during the development of pre-eclampsia starting with a dysregulated trophoblast stem-cell maturation. We provide new targets for potential early disease prevention in order to protect mother and child from increased gestational mortality and morbidity but also from life-long increased cardiovascular disease risk.
Project description:MiRNAs are important posttranscriptional regulators in various physiological processes and their dysregulations have been found in diseases such as infection, imflammation and cancer. Here we compared the miRNA profilings between HBV-producing HepG2.2.15 and its non-HBV maternal HepG2 cell to find potential miRNAs involving in HBV expression. The result indicated that some of the deregulated miRNAs in HepG2.2.15 have been implicated in viral infection and the processes of the immune regulation. HepG2.2.15 vs. HepG2
Project description:We used a whole genome approach to identify major functional gene categories (including xenobiotic transporters and metabolizing enzymes) whose expression depends on gestational age. STUDY DESIGN: We compared gene expression profiles of 1st (45-59 days) and 2nd trimester (109-115 days), and C-section term placentae. RESULTS: In 1st trimester placentae, genes related to cell cycle, DNA, aminoacids and carbohydrate metabolism were significantly overrepresented, while genes related to signal transduction were downregulated. In the organism defense category, we identified genes involved in chemical response, metabolism, and transport. Analysis of signal transduction pathways suggested, and subsequently confirmed independently, that the Wnt pathway was regulated by gestational age. CONCLUSIONS: Our study will serve as a reference database to gain insight into the regulation of gene expression in the developing placentae and, thus, allow comparisons with placentae from complicated pregnancies such as those in women experiencing gestational diabetes, pre-eclampsia and teratogenic sequelae. Experiment Overall Design: Comparison of plaental gene expression profiles of three gestational stages (1st trimester (45-59 days), 2nd trimester (109-115 days), and C-section term placentae), with four replicates each,
Project description:Hypertensive disorders in pregnancy, of which the multisystem syndrome pre-eclampsia is the most severe, leading to preterm delivery, maternal mortality, and life-long complications. To elucidate early disease dynamics, we present the first spatio-temporal study comparing single-nuclei transcriptomes of human preterm pre-eclamptic placentae and healthy controls, contextualizing this in a comprehensive study including early and late gestational placentae. This study includes early placentae samples from the fetal part (villi; n=10), maternal part (Decidua; n=3), late placentae samples from healthy pregnancies, villi (n=6), decidua (n=4), and late placentae samples from early-onset preeclamptic pregnancies, villi (n=5) and decidua (n=5).
Project description:To explore the roles of essential miRNAs in regulating self-renewal of breast cancer stem cells (BCSCs), which initiate from mammary epithelial stem cells (MaSCs). CD44+CD24-/low cells and MUC1-ESA+ cells were isolated by fluorescence-activated cell sorting (FACS) from breast cancer cell line MCF-7 and normal mammary epithelial cell line MCF-10A, and were verified as BCSCs and MaSCs by clonogenic assay and multipotential differentiation experiment in 2-dimensional (2-D) and 3-D cultures, respectively. Using microarray containing oligonucleotides corresponding to 509 miRNAs from human, mouse, and rat genomes. We obtained candidate miRNAs in regulating breast tumorigenesis. One representative miRNA (miR-200c) was proved to regulate stemness of BCSCs and MaSCs in vitro and in vivo by miR-200c agomir transfection. We validated that miR-200c negatively regulated PDCD10, an apoptosis regulator, in BCSCs and MaSCs. Our miRNA microarray includes 509 mature miRNA sequences were downloaded from the miRNA Registry. We designed eight short oligos possessing no homology with all existed RNA sequence and produced their corresponding synthetic miRNAs by in vitro transcription using Ambion’s miRNA Probe Construction kit. All miRNA probe sequences were designed to be complementary to the full-length mature miRNA. The probe sequence were concatenated up to 40 nt and modified with 5 0 -amino-modifier C6. Oligonucleotide probes were synthesized and dissolved in EasyArray TM spotting solution at 40 mM concentration. Each probe was printed in triplicate using a SmartArray TM microarrayer. Arrays were scanned with a confocal LuxScan TM scanner (CapitalBio Corp.)Significance Analysis of Microarrays(SAM, version 2.1) was performed using two class-unpaired comparison in the SAM procedure.
Project description:We used a whole genome approach to identify major functional gene categories (including xenobiotic transporters and metabolizing enzymes) whose expression depends on gestational age. STUDY DESIGN: We compared gene expression profiles of 1st (45-59 days) and 2nd trimester (109-115 days), and C-section term placentae. RESULTS: In 1st trimester placentae, genes related to cell cycle, DNA, aminoacids and carbohydrate metabolism were significantly overrepresented, while genes related to signal transduction were downregulated. In the organism defense category, we identified genes involved in chemical response, metabolism, and transport. Analysis of signal transduction pathways suggested, and subsequently confirmed independently, that the Wnt pathway was regulated by gestational age. CONCLUSIONS: Our study will serve as a reference database to gain insight into the regulation of gene expression in the developing placentae and, thus, allow comparisons with placentae from complicated pregnancies such as those in women experiencing gestational diabetes, pre-eclampsia and teratogenic sequelae. Keywords: time series
Project description:Background: Chronic obstructive pulmonary disease (COPD) is a heterogeneous disease characterized by varying degrees of emphysematous lung destruction and small airway disease, each with distinct effects on clinical outcomes. There is little known about how microRNAs contribute specifically to the emphysema phenotype. We examined how microRNA expression is altered with regional emphysema severity within the lung and how these microRNAs regulate disease-associated gene-expression networks. Results: We profiled microRNAs in different regions in the lung with varying degrees of emphysema from 6 smokers with COPD and 2 controls (8 regions x 8 lungs = 64 samples). 63 microRNAs (p<0.05) were altered with regional emphysema severity as quantified by mean linear intercept (Lm). MicroRNA and gene expression data were then integrated in the same samples. A subset of microRNAs, including miR-638, miR-30c, and miR-181d, correlated with many of their predicted gene targets, suggesting a role in regulating the gene networks that underlie emphysematous lung destruction. Modulating miR-638 expression in primary human lung fibroblasts recapitulated the alterations in its targeted gene-expression network associated with emphysema progression. Pathway analysis revealed that genes involved in oxidative stress and accelerated aging were affected by miR-638 knock-down in fibroblasts. Many miR-638 gene targets in these pathways were amongst those negatively correlated with miR-638 expression in emphysematous lung tissue. Conclusions: Our findings demonstrate that microRNAs are altered with regional emphysema severity and modulate disease-associated gene expression networks. Furthermore, miR-638 may regulate gene expression associated with the oxidative stress response and aging in emphysematous lung tissue and fibroblasts. Paired samples were obtained from 8 regions at regular intervals between the apex and base of each explanted lung from six patients with severe COPD and two donors. The degree of emphysematous destruction was quantified in one sample from each region by mean linear intercept (Lm), while microRNA expression was profiled in the adjacent sample from the same region using the NCode Version 3 microRNA microarrays (Invitrogen, Carlsbad, CA). After quality control 60 samples were used for the analysis.