scholarly journals LncRNAs Function as Critical Metabolic Regulators in yak Liver by Integrative Transcriptome Analysis

2020 ◽  
Author(s):  
Wei Xia ◽  
Fang Fu ◽  
Li Wang ◽  
Xiaolin Luo ◽  
Jiuqiang Guan
Keyword(s):  
Tibetan Plateau ◽  
Correlation Analysis ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Rna Seq ◽  
Qrt Pcr ◽  
The People ◽  
Plateau Area ◽  
Metabolic Regulators

Abstract Background: The yak (Bos grunniens) is a crucial resource to supply meat and milk to the people localized in Qinghai-Tibetan plateau area. To identify lncRNAs regulating metabolism in yak, this work adopted transcriptome method to simultaneously profile mRNAs and lncRNAs of liver in yak under three representative age (LD: Liver 1 Day, LM: Liver 15 Months, LY: Liver 5 Years) conditions.Result: Of 288 differentially expressed lncRNAs, function-oriented selection yield 88 regulated metabolically related lncRNAs that were differentially expressed at least two age conditions. These lncRNAs predicted by lncRNA-mRNA correlation analysis to function in various aspects of metabolism. Selected regulations of liver metabolically related lncRNAs were further verified by qRT-PCR.Conclusion: Combining high throughput RNA-seq screening screens, bioinformatics predictions, lncRNA-mRNA correlation analysis and qRT-PCR analysis, this study supports that a class of lncRNAs function as important metabolic regulators and establishes a foundation for further investigating the role of lncRNAs in yak.

scholarly journals Integrative Transcriptome Analysis Define Novel lncRNA Potential Regulation Function through SLC22A7 in Yak Liver

2021 ◽  
Author(s):  
Wei Xia ◽  
Fang Fu ◽  
Li Wang ◽  
Xiaolin Luo ◽  
Jiuqiang Guan
Keyword(s):  
Correlation Analysis ◽  
Organic Anion ◽  
Organic Anion Transporter ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
The People ◽  
Anion Transporter ◽  
Plateau Area ◽  
Regulation Function

Abstract Background: The yak (Bos grunniens) is a crucial resource to supply meat and milk to the people localized in Qinghai-Tibetan plateau area. To identify lncRNAs regulating metabolism in yak, this work adopted transcriptome method to simultaneously profile mRNAs and lncRNAs of liver in yak under three representative age (LD: Liver 1 Day, LM: Liver 15 Months, LY: Liver 5 Years) conditions.Result: Of 288 differentially expressed lncRNAs, function-oriented selection yield 88 regulated metabolically related lncRNAs that were differentially expressed at least two age conditions. These lncRNAs predicted by lncRNA-mRNA correlation analysis to function in various aspects of metabolism. Selected regulations of liver metabolically related lncRNAs were further verified by qRT-PCR. Furthermore, one novel LncRNA were selected to validate its function and result showed it potentially regulated SLC22A7, a well-known organic anion transporter.Conclusion: Combining high throughput RNA-seq screening screens, bioinformatics predictions, lncRNA-mRNA correlation analysis and qRT-PCR analysis, this study supports that a class of lncRNAs function as important metabolic regulators and establishes a foundation for further investigating the role of lncRNAs in yak.

scholarly journals Integrative Transcriptome Analyses of Metabolic Responses in Yak Liver Define Putative Pivotal LncRNAs

2020 ◽  
Author(s):  
Li Wang ◽  
Wei Xia ◽  
Fang Fu ◽  
Xiaolin Luo ◽  
Jiuqiang Guan
Keyword(s):  
Metabolic Responses ◽  
Qrt Pcr ◽  
The People ◽  
Genome Wide ◽  
Plateau Area ◽  
Physiological Homeostasis ◽  
Transcriptome Analyses ◽  
Metabolic Regulators ◽  
Integrative Transcriptome Analyses

Abstract Background The yak (Bos grunniens) is a crucial resource to supply meat and milk to the people localized in Tibetan plateau area. To systemically identify lncRNAs regulating metabolism in yak, we performed transcriptome analyses to simultaneously profile mRNAs and lncRNAs of liver in yak under three representative age (LD: Liver 1 Day, LM: Liver 15 Month, LY: Liver 5 Year) conditions. Result Of 288 differentially regulated lncRNAs, function-oriented filters yield 88 regulated metabolically related lncRNAs that are differentially expressed at least two age conditions. These lncRNAs predicted by lncRNA-mRNA correlation analyses to function in diverse aspects of metabolism. Specific regulations of liver metabolically related lncRNAs were further defined by qRT-PCR. Conclusion Combining genome-wide screens, bioinformatics function predictions, and qRT-PCR analyses, this study supports that a class of lncRNAs function as important metabolic regulators and establishes a framework for systemically investigating the role of lncRNAs in physiological homeostasis of yak.

scholarly journals ITRAQ-based quantitative proteomic analysis of Fusarium moniliforme (Fusarium verticillioides) in response to Phloridzin inducers

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Rong Zhang ◽  
Weitao Jiang ◽  
Xin Liu ◽  
Yanan Duan ◽  
Li Xiang ◽  
...  
Keyword(s):  
Fusarium Moniliforme ◽  
Abiotic Factors ◽  
Fusarium Verticillioides ◽  
Protein Profile ◽  
Sugar Metabolism ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Differentially Expressed Proteins ◽  
Apple Replant Disease ◽  
Qrt Pcr

Abstract Background Apple replant disease (ARD) has been reported from all major fruit-growing regions of the world, and is often caused by biotic factors (pathogen fungi) and abiotic factors (phenolic compounds). In order to clarify the proteomic differences of Fusarium moniliforme under the action of phloridzin, and to explore the potential mechanism of F. moniliforme as the pathogen of ARD, the role of Fusarium spp. in ARD was further clarified. Methods In this paper, the quantitative proteomics method iTRAQ analysis technology was used to analyze the proteomic differences of F. moniliforme before and after phloridzin treatment. The differentially expressed protein was validated by qRT-PCR analysis. Results A total of 4535 proteins were detected, and 293 proteins were found with more than 1.2 times (P< 0.05) differences. In-depth data analysis revealed that 59 proteins were found with more than 1.5 times (P< 0.05) differences, and most proteins were consistent with the result of qRT-PCR. Differentially expressed proteins were influenced a variety of cellular processes, particularly metabolic processes. Among these metabolic pathways, a total of 8 significantly enriched KEGG pathways were identified with at least 2 affiliated proteins with different abundance in conidia and mycelium. Functional pathway analysis indicated that up-regulated proteins were mainly distributed in amino sugar, nucleotide sugar metabolism, glycolysis/ gluconeogenesis and phagosome pathways. Conclusions This study is the first to perform quantitative proteomic investigation by iTRAQ labeling and LC-MS/MS to identify differentially expressed proteins in F. moniliforme under phloridzin conditions. The results confirmed that F. moniliforme presented a unique protein profile that indicated the adaptive mechanisms of this species to phloridzin environments. The results deepened our understanding of the proteome in F. moniliforme in response to phloridzin inducers and provide a basis for further exploration for improving the efficiency of the fungi as biocontrol agents to control ARD.

scholarly journals Circular RNA Expression Profiling Identifies Prostate Cancer- Specific circRNAs in Prostate Cancer

2018 ◽  
Vol 50 (5) ◽  
pp. 1903-1915 ◽  
Author(s):  
Qianlin Xia ◽  
Tao Ding ◽  
Guihong Zhang ◽  
Zehuan Li ◽  
Ling Zeng ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Polymerase Chain Reaction Analysis ◽  
Circular Rna ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
High Morbidity ◽  
Qrt Pcr ◽  
Diagnosis And Prognosis ◽  
Novel Biomarkers ◽  
Rna Expression Profiling

Background/Aims: Prostate cancer (PCa) is one of the main cancers that damage males’ health severely with high morbidity and mortality, but there is still no ideal molecular marker for the diagnosis and prognosis of prostate cancer. Methods: To determine whether the differentially expressed circRNAs in prostate cancer can serve as novel biomarkers for prostate cancer diagnosis, we screened differentially expressed circRNAs using SBC-ceRNA array in 4 pairs of prostate tumor and paracancerous tissues. A circRNA-miRNA-mRNA regulatory network for the differential circRNAs and their host genes was constructed by Cytoscape3.5.1 software. Quantitative real-time polymerase chain reaction analysis (qRT-PCR) was performed to confirm the microarray data. Results: We found 1021 differentially expressed circRNAs in PCa tumor using SBC-ceRNA array and confirmed the expression of circ_0057558, circ_0062019 and SLC19A1 in PCa cell lines and tumor tissues through qRT-PCR analysis. We demonstrated that combination of PSA level and two differentially expressed circRNAs showed significantly increased AUC, sensitivity and specificity (0.938, 84.5% and 90.9%, respectively) than PSA alone (AUC of serum PSA was 0.854). Moreover, circ_0057558 was correlated positively with total cholesterol. The functional network of circRNA-miRNA-mRNA analysis showed that circ_0057558 and circ_0034467 regulated miR-6884, and circ_0062019 and circ_0060325 regulated miR-5008. Conclusion: Our results demonstrated that differentially expressed circRNAs (circ_0062019 and circ_0057558) and host gene SLC19A1 of circ_0062019 could be used as potential novel biomarkers for prostate cancer.

scholarly journals Chorionic Villous Mesenchymal Stem Cells Derived Exosomes Deliver miR-135b-5p to Trophoblasts, Promoting their Proliferation and Invasion by Targeting TXNIP via β-Catenin Pathway

2020 ◽  
Author(s):  
Yijing Chu ◽  
Yan Zhang ◽  
Guoqiang Gao ◽  
Jun Zhou ◽  
Yang Lv ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Tissue Injury ◽  
Luciferase Reporter ◽  
Rna Seq ◽  
Qrt Pcr ◽  
Reporter Assays ◽  
Pcr Assays ◽  
Proliferation And Invasion

Abstract Background: Human chorionic villous mesenchymal stem cells (CV-MSCs) are found to be a promising and effective treatment for tissue injury. Trophoblast dysfunction during pregnancies is significantly involved in the pathogenesis of preeclampsia (PE). This work was to understand how CV-MSCs regulated trophoblast function. Methods: In this study, we treated trophoblasts with CV-MSC-derived exosomes and RNA-seq analysis was used to understand the changes in trophoblasts. We examined the levels of TXNIP and β-catenin in trophoblasts by immunohistochemistry, western blot and qRT-PCR assays. Luciferase reporter assays and qRT-PCR assays were used to understand the role of miR135b-5p in the effects of CV-MSC-derived exosomes. The growth and invasion of trophoblasts was evaluated with the CCK-8 and transwell assays. Results: The treatment markedly enhanced the trophoblast proliferation and invasion. Furthermore, a significant decrease of TXNIP expression and inactivation of the β-catenin pathway in CV-MSCs exosomes-treated trophoblasts was observed. Consistent with these findings, TXNIP inhibition exhibited the same effect of promoting trophoblast proliferation and invasion as induced by CV-MSC-derived exosomes, also with the accompaniment of inactivation of β-catenin pathway. In addition, overexpression of TXNIP activated the β-catenin pathway in trophoblasts, and reduced the proliferation and invasion of trophoblasts. Importantly, miR135b-5p was found to be highly expressed in CV-MSC exosomes and interact with TXNIP. The miR-135b-5p overexpression significantly elevated the proliferation and invasion of trophoblasts, which could be attenuated by TXNIP overexpression. Conclusion: Our results suggest that TXNIP-dependent β-catenin pathway inactivation mediated by miR135b-5p which is delivered by CV-MSC-derived exosomes could promote the proliferation and invasion of trophoblasts.

scholarly journals Transcriptomic Analysis of Dark-Induced Senescence in Bermudagrass (Cynodon dactylon)

Plants ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 614
Author(s):  
Jibiao Fan ◽  
Yanhong Lou ◽  
Haiyan Shi ◽  
Liang Chen ◽  
Liwen Cao
Keyword(s):  
Leaf Senescence ◽  
Plant Hormone ◽  
Bioinformatics Analysis ◽  
Cynodon Dactylon ◽  
Differential Expression Analysis ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Aesthetic Quality ◽  
Qrt Pcr

Leaf senescence induced by prolonged light deficiency is inevitable whenever turfgrass is cultivated in forests, and this negatively influences the survival and aesthetic quality of the turfgrass. However, the mechanism underlying dark-induced senescence in turfgrass remained obscure. In this study, RNA sequencing was performed to analyze how genes were regulated in response to dark-induced leaf senescence in bermudagrass. A total of 159,207 unigenes were obtained with a mean length of 948 bp. The differential expression analysis showed that a total of 59,062 genes, including 52,382 up-regulated genes and 6680 down-regulated genes were found to be differentially expressed between control leaves and senescent leaves induced by darkness. Subsequent bioinformatics analysis showed that these differentially expressed genes (DEGs) were mainly related to plant hormone (ethylene, abscisic acid, jasmonic acid, auxin, cytokinin, gibberellin, and brassinosteroid) signal transduction, N-glycan biosynthesis, and protein processing in the endoplasmic reticulum. In addition, transcription factors, such as WRKY, NAC, HSF, and bHLH families were also responsive to dark-induced leaf senescence in bermudagrass. Finally, qRT-PCR analysis of six randomly selected DEGs validated the accuracy of sequencing results. Taken together, our results provide basic information of how genes respond to darkness, and contribute to the understanding of comprehensive mechanisms of dark-induced leaf senescence in turfgrass.

scholarly journals Small RNA sequencing evaluation of renal microRNA biomarkers in dogs with X-linked hereditary nephropathy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Candice P. Chu ◽  
Shiguang Liu ◽  
Wenping Song ◽  
Ethan Y. Xu ◽  
Mary B. Nabity
Keyword(s):  
Small Rna ◽  
Target Genes ◽  
Differentially Expressed ◽  
Small Rna Sequencing ◽  
Rna Seq ◽  
Ckd Progression ◽  
Critical Pathways ◽  
Qrt Pcr ◽  
Hereditary Nephropathy ◽  
Differentially Expressed Mirnas

AbstractDogs with X-linked hereditary nephropathy (XLHN) are an animal model for Alport syndrome in humans and progressive chronic kidney disease (CKD). Using mRNA sequencing (mRNA-seq), we have characterized the gene expression profile affecting the progression of XLHN; however, the microRNA (miRNA, miR) expression remains unknown. With small RNA-seq and quantitative RT-PCR (qRT-PCR), we used 3 small RNA-seq analysis tools (QIAGEN OmicSoft Studio, miRDeep2, and CPSS 2.0) to profile differentially expressed renal miRNAs, top-ranked miRNA target genes, and enriched biological processes and pathways in CKD progression. Twenty-three kidney biopsies were collected from 5 dogs with XLHN and 4 age-matched, unaffected littermates at 3 clinical time points (T1: onset of proteinuria, T2: onset of azotemia, and T3: advanced azotemia). We identified up to 23 differentially expressed miRNAs at each clinical time point. Five miRNAs (miR-21, miR-146b, miR-802, miR-142, miR-147) were consistently upregulated in affected dogs. We identified miR-186 and miR-26b as effective reference miRNAs for qRT-PCR. This study applied small RNA-seq to identify differentially expressed miRNAs that might regulate critical pathways contributing to CKD progression in dogs with XLHN.

scholarly journals Human Epicardial Fat Expresses Glucagon-Like Peptide 1 and 2 Receptors Genes

2017 ◽  
Vol 49 (08) ◽  
pp. 625-630 ◽  
Author(s):  
Gianluca Iacobellis ◽  
Vladimir Camarena ◽  
David Sant ◽  
Gaofeng Wang
Keyword(s):  
Subcutaneous Fat ◽  
Metabolic Effects ◽  
Pcr Analysis ◽  
Rna Seq ◽  
Elective Cardiac Surgery ◽  
Qrt Pcr ◽  
Glucagon Like Peptide ◽  
Glucagon Like Peptide 1 ◽  
Artery Disease ◽  
Successful Amplification

AbstractEpicardial adipose tissue (EAT) is an easily measurable visceral fat of the heart with unique anatomy, functionality, and transcriptome. EAT can serve as a therapeutic target for pharmaceutical agents targeting the fat. Glucagon-like peptide-1 (GLP-1) and GLP-2 analogues are newer drugs showing beneficial cardiovascular and metabolic effects. Whether EAT expresses GLP- 1 and 2 receptors (GLP-1R and GLP-2R) is unknown. RNA-seq analysis and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to evaluate the presence of GLP-1R and GLP-2R in EAT and subcutaneous fat (SAT) obtained from 8 subjects with coronary artery disease and type 2 diabetes mellitus undergoing elective cardiac surgery. Immunofluorescence was also performed on EAT and SAT samples using Mab3f52 against GLP-1R. Our RNA-sequencing (RNA-seq) analysis showed that EAT expresses both GLP-1R and GLP-2R genes. qRT-PCR analysis confirmed that GLP-1R expression was low but detected by 2 different sets of intron-spanning primers. GLP-2R expression was detected in all patients and was found to be 5-fold higher than GLP-1R. The combination of accurately spliced reads from RNA-seq and successful amplification using intron-spanning primers indicates that both GLP-1R and GLP-2R are expressed in EAT. Immunofluorescence clearly showed that GLP-1R is present and more abundant in EAT than SAT. This is the first time that human EAT is found to express both GLP-1R and GLP-2R genes. Pharmacologically targeting EAT may induce beneficial cardiovascular and metabolic effects.

scholarly journals Identification of Differentially Expressed MicroRNAs in Osteosarcoma

Sarcoma ◽  
2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
Rishi R. Lulla ◽  
Fabricio F. Costa ◽  
Jared M. Bischof ◽  
Pauline M. Chou ◽  
Maria de F. Bonaldo ◽  
...  
Keyword(s):  
High Throughput ◽  
Mirna Expression ◽  
Expression Profiling ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Osteosarcoma Cell ◽  
Human Osteoblasts ◽  
Normal Human ◽  
Mirna Expression Profiling

A limited number of reports have investigated the role of microRNAs in osteosarcoma. In this study, we performed miRNA expression profiling of osteosarcoma cell lines, tumor samples, and normal human osteoblasts. Twenty-two differentially expressed microRNAs were identified using high throughput real-time PCR analysis, and 4 (miR-135b, miR-150, miR-542-5p, and miR-652) were confirmed and validated in a different group of tumors. Both miR-135b and miR-150 have been previously shown to be important in cancer. We hypothesize that dysregulation of differentially expressed microRNAs may contribute to tumorigenesis. They might also represent molecular biomarkers or targets for drug development in osteosarcoma.

Identification of Differentially Expressed and Spliced Genes with RNA Sequencing Analysis of CLL Specimens

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4582-4582
Author(s):  
Wei Liao ◽  
Gwen Jordaan ◽  
Artur Jaroszewicz ◽  
Matteo Pellegrini ◽  
Sanjai Sharma
Keyword(s):  
B Cells ◽  
Rna Sequencing ◽  
Differentially Expressed Genes ◽  
Fold Change ◽  
Differentially Expressed ◽  
Pcr Analysis ◽  
Sequencing Analysis ◽  
Rna Seq ◽  
Mrna Isoforms ◽  
Core Set

Abstract Abstract 4582 High throughput sequencing of cellular mRNA provides a comprehensive analysis of the transcriptome. Besides identifying differentially expressed genes in different cell types, it also provides information of mRNA isoforms and splicing alterations. We have analyzed two CLL specimens and a normal peripheral blood B cells mRNA by this approach and performed data analysis to identify differentially expressed and spliced genes. The result showed CLLs specimens express approximately 40% more transcripts compared to normal B cells. The FPKM data (fragment per kilobase of exon per million) revealed a higher transcript expression on chromosome 12 in CLL#1 indicating the presence of trisomy 12, which was confirmed by fluorescent in-situ hybridization assay. With a two-fold change in FPKM as a cutoff and a p value cutoff of 0.05 as compared to the normal B cell control, 415 genes and 174 genes in CLL#1 and 676 and 235 genes in CLL#2 were up and downregulated or differentially expressed. In these two CLL specimens, 45% to 75% of differentially expressed genes are common to both the CLL specimens indicating that genetically disparate CLL specimens have a high percentage of a core set of genes that are potentially important for CLL biology. Selected differentially expressed genes with increased expression (selectin P ligand, SELPLG, and adhesion molecule interacts with CXADR antigen 1, AMICA) and decreased (Fos, Jun, CD69 and Rhob) expression based on the FPKM from RNA-sequencing data were also analyzed in additional CLL specimens by real time PCR analysis. The expression data from RNA-seq closely matches the fold-change in expression as measured by RT-PCR analysis and confirms the validity of the RNA-seq analysis. Interestingly, Fos was identified as one of the most downregulated gene in CLL. Using the Cufflinks and Cuffdiff software, the splicing patterns of genes in CLL specimens and normal B cells were analyzed. Approximately, 1100 to 1250 genes in the two CLL specimens were significantly differentially spliced as compared to normal B cells. In this analysis as well, there is a core set of 800 common genes which are differentially spliced in the two CLL specimens. The RNA-sequencing analysis accurately identifies differentially expressed novel genes and splicing variations that will help us understand the biology of CLL. Disclosures: No relevant conflicts of interest to declare.

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