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 Effect of miR-27b-5p on apoptosis of human vascular endothelial cells induced by simulated microgravity

APOPTOSIS ◽  
2019 ◽  
Vol 25 (1-2) ◽  
pp. 73-91 ◽  
Author(s):  
Yi-Kai Pan ◽  
Cheng-Fei Li ◽  
Yuan Gao ◽  
Yong-Chun Wang ◽  
Xi-Qing Sun
Keyword(s):  
Endothelial Cells ◽  
Western Blot ◽  
Target Gene ◽  
Simulated Microgravity ◽  
Target Genes ◽  
Expression Profiles ◽  
Differentially Expressed ◽  
Luciferase Assay ◽  
Qrt Pcr ◽  
Differentially Expressed Mirnas

AbstractWeightlessness-induced cardiovascular dysfunction can lead to physiological and pathological consequences. It has been shown that spaceflight or simulated microgravity can alter expression profiles of some microRNAs (miRNAs). Here, we attempt to identify the role of miRNAs in human umbilical vein endothelial cells (HUVECs) apoptosis under simulated microgravity. RNA-sequencing and quantitative real-time PCR (qRT-PCR) assays were used to identify differentially expressed miRNAs in HUVECs under simulated microgravity. Then we obtained the target genes of these miRNAs through target analysis software. Moreover, GO and KEGG enrichment analysis were performed. The effects of these miRNAs on HUVECs apoptosis were evaluated by flow cytometry, Western blot and Hoechst staining. Furthermore, we obtained the target gene of miR-27b-5p by luciferase assay, qRT-PCR and Western blot. Finally, we investigated the relationship between this target gene and miR-27b-5p in HUVECs apoptosis under normal gravity or simulated microgravity. We found 29 differentially expressed miRNAs in HUVECs under simulated microgravity. Of them, the expressions of 3 miRNAs were validated by qRT-PCR. We demonstrated that miR-27b-5p affected HUVECs apoptosis by inhibiting zinc fingers and homeoboxes 1 (ZHX1). Our results reported here demonstrate for the first time that simulated microgravity can alter the expression of some miRNAs in HUVECs and miR-27b-5p may protect HUVECs from apoptosis under simulated microgravity by targeting ZHX1.

scholarly journals Identification and study of differentially expressed miRNAs in aged NAFLD rats based on high-throughput sequencing

2020 ◽  
Vol 19 (3) ◽  
pp. 302-312 ◽  
Author(s):  
Ying Zhang ◽  
Danni Xiang ◽  
Xiaona Hu ◽  
Qingwei Ruan ◽  
Lina Wang ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Sequencing ◽  
Differentially Expressed ◽  
Differentially Expressed Mirnas

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 &lt; 0.05, Fold change &gt;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 High-Throughput Sequencing Reveals the Differential MicroRNA Expression Profiles of Human Gastric Cancer SGC7901 Cell Xenograft Nude Mouse Models Treated with Traditional Chinese Medicine Si Jun Zi Tang Decoction

2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Junfu Guo ◽  
Xiangnan Li ◽  
Lanying Miao ◽  
Hongwei Sun ◽  
Xia Gao ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
High Throughput Sequencing ◽  
Target Genes ◽  
Expression Profiles ◽  
Differentially Expressed ◽  
Human Gastric Cancer ◽  
Model Group ◽  
Sgc7901 Cell ◽  
Qrt Pcr ◽  
Differentially Expressed Mirnas

Objective. The present study aimed to investigate the potential mechanism underlying the antitumor effect of Si Jun Zi Tang (SJZT) decoction on gastric cancer. Methods. Twelve human gastric cancer SGC7901 cell xenograft nude mouse models were established. The mice were randomly divided into the Model group and SJZT group. SJZT exerted significant antitumor effects after 21 days of decoction administration. High-throughput sequencing was used to analyze the microRNA (miRNA) expression profiles of tumor tissues. Bioinformatics analysis was performed to provide further information regarding the differentially expressed miRNAs. Five representative differentially expressed miRNAs and four predicted target genes were further validated using quantitative real-time reverse transcription PCR (qRT-PCR). Results. We identified 33 miRNAs that were differentially expressed in the SJZT group compared with the Model group. Among them, 32 miRNAs were upregulated and 1 miRNA was downregulated. Bioinformatic analysis showed that most of miRNAs acted as tumor suppressors and their target genes participated in multiple signaling pathways, including the PI3K/Akt signaling pathway, microRNAs in cancer, and Wnt signaling pathway. The qRT-PCR result confirmed that miR-223-3p, miR-205-5p, miR-147b-3p, and miR-223-5p were overexpressed and their respective paired target genes FUT9, POU2F1, MUC4, and RAB14 mRNA were obviously downregulated in the SJZT group compared with those in the Model group. Network analysis revealed that miR-223-3p and miR-205-5p shared two targets POU2F1 (encoding POU class 2 homeobox 1) and FUT9 (encoding fucosyltransferase 9), suggesting they have a common role in certain pathways. Conclusion. This study provided novel insights into the anticancer mechanism of SJZT against gastric cancer, which might be partly related to the modulation of miRNA expression and their target pathways in tumors.

Stretch-Induced Lncrna SNHG8 Inhibits Osteogenic Differentiation by Regulating EZH2 in Hpdlscs

2021 ◽  
Author(s):  
Zijie Zhang ◽  
Qin He ◽  
Xiaolu Zhao ◽  
Xiaoyu Li ◽  
Fulan Wei
Keyword(s):  
Alkaline Phosphatase ◽  
Western Blot ◽  
Osteogenic Differentiation ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Tooth Movement ◽  
Alizarin Red ◽  
Qrt Pcr ◽  
Osteogenic Induction

Abstract Background: Periodontal ligament stem cells (PDLSCs) are important for the remodeling of the alveolar bone while tooth moving. However, the effect of long non-coding RNA (lncRNA) on osteogenic differentiation of PDLSCs under mechanical force remains unclear.Methods: In this study, we compared stretched and non-stretched PDLSCs by high-throughput sequencing. The verification and selection of lncRNAs were achieved by quantitative reverse transcription polymerase chain reaction (qRT-PCR). PDLSCs osteogenic differentiation potentials were assessed by alkaline phosphatase (ALP) staining, Alizarin Red staining, qRT-PCR, and western blot. The application of mechanical force used Flexcell-FX-6000-Tension System in vitro, and constructing rats’ tooth movement model in vivo. To verify the osteogenic regulation ability of small nucleolar RNA host gene 8 (SNHG8), PDLSCs were stretched or applied osteogenic induction after been infected by lentivirus. RNA fluorescence in situ hybridization, isolation of nuclear and cytoplasmic RNA, qRT-PCR and western blot were performed to locate SNHG8. Western blot and qRT-PCR to find the relationship between enhancer of zeste homolog 2 (EZH2) and SNHG8.Results: Our results demonstrated that among lncRNAs altered screened by high-throughput sequencing, the expression level of SNHG8 steadily decreased after being stretched. Analysis of mRNA expression and protein levels revealed an upregulation of ALP and RUNX2, ALP and Alizarin Red staining showed more obvious alkaline phosphatase and more mineralized nodules in SNHG8 knockdown PDLSCs. In vivo experiments showed lower expression of the homologous gene of SNHG8 after tooth movement, and better ability of ectopic osteogenesis after knockdown SNHG8. The verification of SNHG8’s nuclear location led us to infer that SNHG8 may interact with EZH2. The qRT-PCR and western blot results disclosed EZH2 expression reduced along with the knockdown of SNHG8. Furthermore, knockdown of EZH2 lead to PDLSCs’ osteogenic differentiation ability increasing under osteogenic induction according to the mRNA level of ALP and RUNX2 accompanied by ALP and Alizarin Red staining results.Conclusion: In general, our study confirmed that mechanically sensitive lncRNA SNHG8 can influence the osteogenic differentiation of PDLSCs through epigenetic pathways without directly encoding protein, which provides solid evidence for the regulation by non-coding genes.

scholarly journals The effects of the Xijiao Dihuang decoction combined with Yinqiao powder on miRNA-mRNA profiles in mice infected with influenza a virus

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Ke Li ◽  
Xiaoming Chen ◽  
Jing Zhong ◽  
Hehe Ye ◽  
Shujing Zhang ◽  
...  
Keyword(s):  
Influenza A ◽  
High Throughput Sequencing ◽  
Enrichment Analysis ◽  
Differentially Expressed ◽  
Kegg Database ◽  
Qrt Pcr ◽  
Go Enrichment ◽  
Differentially Expressed Mirnas ◽  
Therapeutic Mechanisms ◽  
Go Enrichment Analysis

Abstract Background MicroRNAs (miRNAs) play vital roles in acute inflammatory and antiviral responses during influenza A virus (IAV) infection. The Xijiao Dihuang decoction combined with Yinqiao powder (XDY) is applied to remedy viral pneumonia in China and its therapeutic efficacy in pneumonic mice challenged with IAV was demonstrated; however, the underlying mechanisms remain elusive. Thus, this study aimed to explore the miRNA-mRNA profiles in the lungs of IAV-infected mice and investigate the therapeutic mechanisms of XDY involving miRNAs and associated pathways. Methods We detected the cellular miRNA contents in the lungs of mice treated with XDY (23 g/kg/d) for A/FM/1/47 (H1N1) (FM1) infection at 4 days postinoculation (dpi) and 7 dpi. MiRNA and mRNA high-throughput sequencing analyses, and miRNA and mRNA qRT-PCR analyses were used to detect and verify the relevant miRNAs and mRNAs. Conjoint analysis, GO enrichment analysis, and KEGG database analysis were applied to identify the miRNA-mRNA regulatory relationships. Results The quantities of differentially expressed miRNAs and mRNAs were upregulated over time. The data showed that 104 miRNAs and 3485 mRNAs were differentially expressed after challenge with FM1 on day 4, while 191 miRNAs and 6126 mRNAs were differentially expressed on day 7. The GO enrichment analysis and KEGG database data showed that the differentially expressed miRNAs and mRNAs were mainly enriched in JNK activity, MAPK phosphatase activity, and the TLR, Jak-STAT and TNF signalling pathways after treatment of FM1 infection with XDY. Generally, the expression trends of differentially expressed miRNAs and mRNAs based on the qRT-PCR results exhibited good consistency with the results of the high-throughput sequencing analysis. Conclusions MiRNAs and mRNAs were differentially expressed during FM1 infection. The therapeutic mechanisms of XDY in FM1-infected mice, might be related to regulating antiviral immunity and ameliorating excessive inflammatory responses by modulating the expression of dysregulated miRNAs and mRNAs involved in the ERK/JNK-AP-1, and IFN-β/STAT signalling pathways.

scholarly journals miRNA Expression Characterizes Histological Subtypes and Metastasis in Penile Squamous Cell Carcinoma

Cancers ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1480
Author(s):  
Hiresh Ayoubian ◽  
Joana Heinzelmann ◽  
Sebastian Hölters ◽  
Oybek Khalmurzaev ◽  
Alexey Pryalukhin ◽  
...  
Keyword(s):  
Microarray Data ◽  
Expression Patterns ◽  
Hpv Infection ◽  
Differentially Expressed ◽  
Histological Subtypes ◽  
Specific Expression ◽  
Tissue Samples ◽  
Qrt Pcr ◽  
Differentially Expressed Mirnas ◽  
The Impact

Although microRNAs are described as promising biomarkers in many tumor types, little is known about their role in PSCC. Thus, we attempted to identify miRNAs involved in tumor development and metastasis in distinct histological subtypes considering the impact of HPV infection. In a first step, microarray analyses were performed on RNA from formalin-fixed, paraffin-embedded tumor (22), and normal (8) tissue samples. Microarray data were validated for selected miRNAs by qRT-PCR on an enlarged cohort, including 27 tumor and 18 normal tissues. We found 876 significantly differentially expressed miRNAs (p ≤ 0.01) between HPV-positive and HPV-negative tumor samples by microarray analysis. Although no significant differences were detected between normal and tumor tissue in the whole cohort, specific expression patterns occurred in distinct histological subtypes, such as HPV-negative usual PSCC (95 differentially expressed miRNAs, p ≤ 0.05) and HPV-positive basaloid/warty subtypes (247 differentially expressed miRNAs, p ≤ 0.05). Selected miRNAs were confirmed by qRT-PCR. Furthermore, microarray data revealed 118 miRNAs (p ≤ 0.01) that were significantly differentially expressed in metastatic versus non-metastatic usual PSCC. The lower expression levels for miR-137 and miR-328-3p in metastatic usual PSCC were validated by qRT-PCR. The results of this study confirmed that specific miRNAs could serve as potential diagnostic and prognostic markers in single PSCC subtypes and are associated with HPV-dependent pathways.

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