scholarly journals Analysis of LncRNA-mRNA Co-Expression Profiles in Patients With Polycystic Ovary Syndrome: A Pilot Study

2021 ◽  
Vol 12 ◽  
Author(s):  
Xiuhong Sun ◽  
Yishan Liu ◽  
Xinyu Gao ◽  
Mengxuan Du ◽  
Mengge Gao ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
High Throughput ◽  
Peripheral Blood ◽  
High Throughput Sequencing ◽  
Polycystic Ovary ◽  
Differentially Expressed ◽  
Ovary Syndrome ◽  
Qrt Pcr ◽  
Chemokine Signaling ◽  
Chemokine Signaling Pathway

PurposeThis study aimed to investigate the profiles of messenger RNAs (mRNAs) and long noncoding RNAs (lncRNAs) in peripheral blood samples collected from polycystic ovary syndrome (PCOS) patients. In addition, an in-depth bioinformatics analysis regarding the lncRNA-mRNA co-expression network was performed.MethodsHigh-throughput sequencing was used to measure the profiles of mRNAs and lncRNAs expressed in the peripheral blood samples isolated from six patients (three patients with PCOS and three normal women). In addition, five differentially expressed lncRNAs were chosen to validate the results of high-throughput sequencing by quantitative RT-PCR (qRT-PCR). Furthermore, a lncRNA-mRNA co-expression network was constructed using the Cytoscape software.ResultsA total of 14,276 differentially expressed mRNAs and 4,048 differentially expressed lncRNAs were obtained from the RNA-seq analysis of PCOS patients and healthy controls (adjusted q-value < 0.05, Fold change >2.0).The qRT-PCR results were consistent with the data obtained through high-throughput sequencing. Gene ontology (GO) and KEGG pathway analyses showed that the differentially expressed mRNAs were enriched in the chemokine signaling pathway. In addition, the analysis of the lncRNA-mRNA co-expression network of the chemokine signaling pathway showed the involvement of 6 mRNAs and 42 lncRNAs.ConclusionClusters of mRNAs and lncRNAs were aberrantly expressed in the peripheral blood of PCOS patients compared with the controls. In addition, several pairs of lncRNA-mRNAs in the chemokine signaling pathway may be related to PCOS genetically.

scholarly journals Evaluation on the Characteristics of Gut Microbiome in Polycystic Ovary Syndrome Rats Induced by Dihydrotestosterone or Letrozole

2020 ◽  
Author(s):  
Yanhua Zheng ◽  
Hongxia Ma ◽  
Ying Xu ◽  
Chengjie Liang ◽  
Tong Yang
Keyword(s):  
Polycystic Ovary Syndrome ◽  
Gut Microbiota ◽  
Aromatase Inhibitor ◽  
16S Rdna ◽  
High Throughput ◽  
Gut Microbiome ◽  
High Throughput Sequencing ◽  
Polycystic Ovary ◽  
Molecular Ecology ◽  
Ovary Syndrome

Abstract The etiology of polycystic ovary syndrome (PCOS) is unclear. Recent reports indicated that the gut microbiome of PCOS patients and rodents has changed. In this study, we induced the nonaromatizable androgen dihydrotestosterone (DHT) or the aromatase inhibitor letrozole (LET) to induce PCOS model rat to compare the bacterial diversity distribution within and between the two groups. The molecular ecology of the fecal gut microbiota was analyzed by 16S rDNA high-throughput sequencing. Our study found that DHT can reduce the microbial richness in rats. PCoA plots confirmed that DHT group was statistically significantly separated from C group and LET group. At phylum level, DHT led to a decrease in Bacteroidetes as well as an increase in Cyanobacteria, Tenericutes, Actinobacteria, Spirochaetae and Saccharibacteria. At genus level, LEfSe analysis showed that genus of Bifidobacteriales, Vibro, Peptococcus and Turicibacter played roles in the letrozole induced PCOS rats. And Lachnospiraceae_NK4A136_group, Ruminococcus_1, Ruminiclostridium, Anaerotruncus and Anaeroplasma played vital roles in the intestine of DHT induced PCOS rats.

scholarly journals Effects of keratinocyte-derived and fibroblast-derived exosomes on human epidermal melanocytes

2021 ◽  
Vol 0 ◽  
pp. 1-10
Author(s):  
Hai-Xia Shi ◽  
Ru-Zhi Zhang ◽  
Li Xiao ◽  
Li Wang
Keyword(s):  
Signaling Pathway ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Target Genes ◽  
Tyrosinase Activity ◽  
The Other ◽  
Differentially Expressed ◽  
Ras Signaling ◽  
Sequencing Analysis ◽  
Melanin Synthesis

Background: Exosomes have been demonstrated to carry proteins, membrane lipids, mRNAs and microRNAs which can be transferred to surrounding cells and regulate the functions of those recipient cells. Objectives: The objective of the study was to investigate the effects of exosomes released by keratinocytes and fibroblasts on the proliferation, tyrosinase activity and melanogenesis of melanocytes. Methods: Melanocytes, keratinocytes and fibroblasts obtained from human foreskin were cultured and exosomes secreted by keratinocytes and fibroblasts were harvested from the culture supernatants by ultracentrifugation. Each exosome fraction was divided into two parts; one part was subjected to high-throughput sequencing using an Illumina HiSeq sequencer to characterize the microRNA expression profiles, while the other part was labeled with the fluorescent dye PKH67 and was then co-cultivated with epidermal melanocytes. Results: High-throughput sequencing analysis showed 168 differentially expressed microRNA within exosomes derived from keratinocytes and from fibroblasts, 97 of those being up-regulated with the other 71 down-regulated. Gene ontology analysis showed that the target genes responsible for these differentially expressed microRNAs were mainly enriched in the protein-binding region of molecular functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that target genes regulated by differentially expressed microRNA were mainly involved in mitogen-activated protein kinase (MAPK) signaling pathway, Ras signaling pathway, cAMP signaling pathway and Wnt signaling pathway. Keratinocyte-derived exosomes were taken up by melanocytes co-cultured with them and promoted the proliferation, tyrosinase activity and melanin synthesis of those melanocytes. However, fibroblast-derived exosomes had no similar effects on melanocytes. Conclusion: Keratinocyte-derived exosomes but not fibroblast-derived exosomes were taken up by melanocytes in co-culture and significantly stimulated their proliferation, tyrosinase activity and melanin synthesis. Those different effects may be mainly due to the differential expression of microRNAs in exosomes derived from the different types of cells. Limitations: Electron microscopy of the obtained exosomes and in-depth study of apparently differentially expressed microRNAs were not performed.

scholarly journals RNA-sequencing reveals the expression profiles of tsRNAs and their potential carcinogenic role in cholangiocarcinoma

2021 ◽  
Author(s):  
Li-rong Yan ◽  
Ang Wang ◽  
Qian Xu ◽  
Ben-gang Wang
Keyword(s):  
Signaling Pathway ◽  
Rna Sequencing ◽  
High Throughput ◽  
Target Genes ◽  
Expression Profiles ◽  
Biological Effects ◽  
Differentially Expressed ◽  
Rna Seq ◽  
Normal Tissues ◽  
Qrt Pcr

Abstract Background Recently, the incidence of cholangiocarcinoma (CCA) has gradually increased. As CCA has a poor prognosis, the ideal survival rate is scarce for patients. The abnormal expressed tsRNAs may regulate the progression of a variety of tumors, and tsRNAs is expected to become a new diagnostic biomarker of cancer. However, the expression of tsRNAs is obscure and should be elucidated in CCA. Methods High-throughput RNA sequencing technology (RNA-seq) was utilized to determine the overall expression profiles of tsRNAs in 3 pairs CCA and adjacent normal tissues and to screen the tsRNAs that were differentially expressed. The target genes of dysregulated tsRNAs were predicted and the biological effects and potential signaling pathways of these target genes were explored by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to validate 11 differentially expressed tRFs with 12 pairs CCA and adjacent normal tissues. Results High-throughput RNA-seq totally demonstrated 535 dysregulated tsRNAs, of which 241 tsRNAs were upregulated and 294 tsRNAs were downregulated in CCA compared with adjacent normal tissues (|log2 (fold change) |≥1 and P value < 0.05). GO and KEGG enrichment analyses indicated that the target genes of dysregulated tRFs (tRF-34-JJ6RRNLIK898HR, tRF-38-0668K87SERM492V and tRF-39-0668K87SERM492E2) were mainly enriched in the Notch signaling pathway, Hippo signaling pathway, cAMP signaling pathway and in growth hormone synthesis, secretion and action, etc. qRT-PCR result showed that tRF-34-JJ6RRNLIK898HR/tRF-38-0668K87SERM492V/tRF-39-0668K87SERM492E2 was down-regulated (P = 0.021) and tRF-20-LE2WMK81 was up-regulated in CCA (P = 0.033). Conclusion Differentially expressed tRFs in CCA are enriched in many pathways associated with neoplasms, which may impact the tumor progression and have potential to be diagnostic biomarkers and therapeutic targets of CCA.

scholarly journals Differences in the transcriptional profiles of human cumulus cells isolated from MI and MII oocytes of patients with polycystic ovary syndrome

Reproduction ◽  
2013 ◽  
Vol 145 (6) ◽  
pp. 597-608 ◽  
Author(s):  
Xin Huang ◽  
Cuifang Hao ◽  
Xiaofang Shen ◽  
Xiaoyan Liu ◽  
Yinghua Shan ◽  
...  
Keyword(s):  
Polycystic Ovary Syndrome ◽  
Polycystic Ovary ◽  
Expression Profiles ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
Differentially Expressed ◽  
Embryo Viability ◽  
Ovary Syndrome ◽  
Developmental Potential ◽  
Qrt Pcr

Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women. The abnormalities of endocrine and intra-ovarian paracrine interactions may change the microenvironment for oocyte development during the folliculogenesis process and reduce the developmental competence of oocytes in PCOS patients who are suffering from anovulatory infertility and pregnancy loss. In this microenvironment, the cross talk between an oocyte and the surrounding cumulus cells (CCs) is critical for achieving oocyte competence. The aim of our study was to investigate the gene expression profiles of CCs obtained from PCOS patients undergoing IVF cycles in terms of oocyte maturation by using human Genome U133 Plus 2.0 microarrays. A total of 59 genes were differentially expressed in two CC groups. Most of these genes were identified to be involved in one or more of the following pathways: receptor interactions, calcium signaling, metabolism and biosynthesis, focal adhesion, melanogenesis, leukocyte transendothelial migration, Wnt signaling, and type 2 diabetes mellitus. According to the different expression levels in the microarrays and their putative functions, six differentially expressed genes (LHCGR, ANGPTL1, TNIK, GRIN2A, SFRP4, and SOCS3) were selected and analyzed by quantitative RT-PCR (qRT-PCR). The qRT-PCR results were consistent with the microarray data. Moreover, the molecular signatures (LHCGR, TNIK, and SOCS3) were associated with developmental potential from embryo to blastocyst stage and were proposed as biomarkers of embryo viability in PCOS patients. Our results may be clinically important as they offer a new potential strategy for competent oocyte/embryo selection in PCOS patients.

scholarly journals LncRNA and transcriptomic analysis of fetal membrane revealed potential targets involved in oligohydramnios

2019 ◽  
Author(s):  
Yu-hua Ou ◽  
Yu-kun Liu ◽  
Li-qiong Zhu ◽  
Man-qi Chen ◽  
Xiao-chun Yi ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Tissue Growth ◽  
Differentially Expressed ◽  
Fetal Membrane ◽  
Ras Signaling ◽  
Chromosome 2 ◽  
Chemokine Signaling ◽  
Wide Range ◽  
Chemokine Signaling Pathway

Abstract The challenge of oligohydramnios study is multiple causes of oligohydramnios. Long noncoding RNAs (lncRNAs) is a set of RNAs that has been proved to function in multiple biological process. Currently, little is know about their expression and possible role in oligohydramnios. Total RNA was isolated from fetal membranes ressected from oligohydramnios pregnant women (OR) and normal amniotic fluid control (Normal).RNA-sequencing (RNA-seq) obtain that a total of 801 lncRNAs and 367 mRNAs were differentially expressed in OR. Of which, 638 lncRNAs and 189 mRNAs were upregulated, and 163 lncRNAs and 178 mRNAs were downregulated. Of these lncRNAs, 566 of them were intergenic lncRNA, 351 were intronic antisense lncRNA，and 300 natural antisense. The differentially expressed lncRNA were primary located in chromosome 2, 1 and 11. KEGG enrichment pathways revealed the differential expressed mRNAs were enriched in pathway in cancer, Ras signaling pathway, TNF signaling pathway, focal adhesion, and chemokine signaling pathway. The qRT-PCR result confirmed that LINC00515 and RP11-388P9.2 were upregulated in OR. Furthermore, the constructed lncRNA-miRNA-mRNA regulatory network revealed TNR, CFTR, ABCC2, ABCA12, and COL9A2 as the candidate targets of LINC00515 and RP11-388P9.2. A wide range of lncRNAs were alert in OR, in particularly, LINC00515 and RP11-388P9.2 were confirmed to be uprgulated in OR, and their predicted downstream targets were transport and tissue growth and development associated. Further study focused on the role of differential expressed lncRNAs such as LINC00515 and RP11-388P9.2 would provide more insight into the pathophysiology in OR.

scholarly journals MON-207 Identification of Monogenic Causes of Polycystic Ovary Syndrome by High Throughput Sequencing

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Raiane Crespo ◽  
Thais Rocha ◽  
Gustavo Maciel ◽  
Sylvia Hayashida ◽  
Edmund Baracat ◽  
...  
Keyword(s):  
Polycystic Ovary Syndrome ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Polycystic Ovary ◽  
Ovary Syndrome

scholarly journals Up－Regulated miR-224-5p Can Target the Expression of Neuritin in Hearing Loss

2021 ◽  
Author(s):  
Yin Song ◽  
Xue Zhang ◽  
Jiawei Sun ◽  
Lina Li ◽  
Xiaofei Zhang ◽  
...  
Keyword(s):  
Hearing Loss ◽  
Western Blot ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Differentially Expressed ◽  
Qrt Pcr ◽  
Kanamycin Sulfate ◽  
Differentially Expressed Mirnas ◽  
Mouse Hearing ◽  
The Relationship

Abstract Objective: This study screened the differentially expressed miRNAs in the mouse cochlea during hearing loss and explored the relationship between miR-224-5p and Neuritin.Methods: The combination of kanamycin sulfate and furosemide was used to establish a mouse hearing loss model. High-throughput sequencing was used to screen the differentially expressed miRNAs during hearing loss. qRT-PCR was used to identify the expression of differential miRNAs in hearing loss. Western Blot was used to detect the expression of Neuritin protein. Luciferase was used to identify the binding site of miRNA and Neuritin.Results: The expression of miR-224-5p in the mouse cochlea increased during hearing loss (p<0.05). MiR-224-5p mimics can reduce Neuritin protein expression in 293T cells (p<0.05). MiR-224-5p can specifically bind to Neuritin (p<0.05).Conclusion: The expression of miR-224-5p increases in hearing loss and targets the expression of Neuritin

scholarly journals Microarray analysis reveals an inflammatory transcriptomic signature in peripheral blood for sciatica

BMC Neurology ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Yi Wang ◽  
Guogang Dai ◽  
Ling Jiang ◽  
Shichuan Liao ◽  
Jiao Xia
Keyword(s):  
Functional Analysis ◽  
Network Analysis ◽  
Peripheral Blood ◽  
Differentially Expressed ◽  
Healthy Controls ◽  
Toll Like Receptor ◽  
Qrt Pcr ◽  
Transcriptomic Signature ◽  
Transcriptional Changes ◽  
Key Genes

Abstract Background Although the pathology of sciatica has been studied extensively, the transcriptional changes in the peripheral blood caused by sciatica have not been characterized. This study aimed to characterize the peripheral blood transcriptomic signature for sciatica. Methods We used a microarray to identify differentially expressed genes in the peripheral blood of patients with sciatica compared with that of healthy controls, performed a functional analysis to reveal the peripheral blood transcriptomic signature for sciatica, and conducted a network analysis to identify key genes that contribute to the observed transcriptional changes. The expression levels of these key genes were assessed by qRT-PCR. Results We found that 153 genes were differentially expressed in the peripheral blood of patients with sciatica compared with that of healthy controls, and 131 and 22 of these were upregulated and downregulated, respectively. A functional analysis revealed that these differentially expressed genes (DEGs) were strongly enriched for the inflammatory response or immunity. The network analysis revealed that a group of genes, most of which are related to the inflammatory response, played a key role in the dysregulation of these DEGs. These key genes are Toll-like receptor 4, matrix metallopeptidase 9, myeloperoxidase, cathelicidin antimicrobial peptide, resistin and Toll-like receptor 5, and a qRT-PCR analysis validated the higher transcript levels of these key genes in the peripheral blood of patients with sciatica than in that of healthy controls. Conclusion We revealed inflammatory characteristics that serve as a peripheral blood transcriptomic signature for sciatica and identified genes that are essential for mRNA dysregulation in the peripheral blood of patients with sciatica.

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