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Exosomal miRNAs differences analysis between C26 and MC38

ABSTRACT: Exosomal miRNAs differences analysis between C26 and MC38

PROVIDER: PRJNA724681 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14310085_1.fastq.gz	Fastqsanger.gz
	SRR14310085_2.fastq.gz	Fastqsanger.gz
	SRR14310086_1.fastq.gz	Fastqsanger.gz
	SRR14310086_2.fastq.gz	Fastqsanger.gz
	SRR14310087_1.fastq.gz	Fastqsanger.gz

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Exosomal miRNAs differences analysis between C26 and MC38

Project description:MiRNAs contained in C26-derived and MC38-derived exosomes were determined by IlluminaHiSeq platform. High-throughput sequencing was conducted at Shanghai MajorbioBiopharm TechnologyCo., Ltd. (Shanghai, China). In brief, total RNA from exosomes was prepared and quantified with a NanoDrop ND-2000(NanoDrop Technologies).RNA integrity was assessed using a 2100 Bioanalyzer (Agilent Technologies,Santa Clara, CA, USA). A total amount of 3 μg total RNA per sample was used as input material for the small RNA library. Small RNA adapters were then ligated to the 5’ and 3’ ends of total RNA. After cDNA synthesis and amplification, the PCR-amplified fragments were purified from the PAGE gel, and the completed cDNA libraries were quantified by an Agilent 2100 Bioanalyzer. Cluster generation was performed on an IlluminacBot, and sequencing was performed on an IlluminaHiSeq 2000 platform and 50bp single-end reads were generated.The expression level of each miRNA was calculated according to the transcripts per million reads (TPM) method. Significant differently expressed (DE) miRNAs were extracted with |log2FC|>1 and FDR < 0.05 by DEseq2.

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Project description:Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin Lymphoma with complex clinical symptoms. Currently, the diagnosis of DLBCL depends largely on pathologic analysis of biopsy tissue, which is an invasive procedure and often poses some risk to patients. Exosomal miRNAs have emerged as promising noninvasive biomarkers for detecting tumor or monitoring disease progress. We aim to investigate the potential of circulating exosomal miRNAs to assist with diagnosis of DLBCL.In the present research, we used the miRCURY LNA™ microRNA Array to investgate the profiles of differentially expressed miRNAs in the exosomes of peripheral blood serum. As a result, circulating exosomal miRNA profiling yielded a total of 332 differentially expressed miRNAs (157 with up-expression and 175 with down-expression) between the DLBCL patients and healthy controls.

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Project description:To investigate the possibilities of circulating exosomal miRNAs in the early screening and prevention of diabetic retinopathy(DR), and to explore how the exosomal miRNAs functioning in DR. We then performed gene expression profiling analysis using data obtained from small RNA-seq of 3 diabetes mellitus(DM) patients and 3 DR patients.

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Project description:To investigate the differences in miRNA profiles, exosomal and cellular miRNA profiles between hCEp (Human Cervical Epithelia Cells) and Siha cells were generated by deep sequencing, in triplicate, using Illumina GAIIx. qRT–PCR validation was performed using TaqMan and SYBR Green assays

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Project description:We used miRNA-seq and bioinformatics to analyze and annotate the expression profiles exosomal miRNAs in pancreatic cancer cells after radiation, compared with the unirradiated cells. A total of 481 miRNAs were identified, and 284 miRNAs were annotated in miRBase. There were 22 filtered differentially expressed miRNAs (9 for up-regulated and 13 for down-regulated, fold change > 2, p-value < 0.05). This study provides the results of exosomal miRNA change in pancreatic cancer cells after radiation.

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Project description:Objective: The expression pattern of exosomal miRNAs derived from parathyroid tumor is still unknown. In the present work, we aimed to examine the differences on microRNA (miRNA) expression, present in serum exosomes, by comparing parathyroid carcinoma (PC) and parathyroid adenoma (PA). Methods: MiRNA expression profile of serum exosomes, derived from 4 PC patients and 4 PA patients, were analyzed by next-generation sequencing technology. The differential expressions of target miRNAs were further verified in both serum exosomes and tissues of PC/PA patients by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Lastly, receiver operating characteristic (ROC) curves were plotted to investigate the efficiency of target exosomal miRNAs in distinguishing PC patients from PA controls. Results: Multiple differentially expressed miRNAs of serum exosomes were screened out by sequencing. Based on this screening, hsa-miR-146b-5p (p=0.0846), hsa-miR-27a-5p (p=0.0412), hsa-miR-93-5p (p=0.73), hsa-miR-381-3p (p=0.1239) and hsa-miR-134-5p (p=0.0694) were up-regulated in the serum exosomes of PC patients. These results were validated by qPCR, where the trend on differential miRNA expression was consistent with the sequencing results. Specifically, the expression of exosomal hsa-miR-27a-5p was able to clearly distinguish PC patients from PA controls, and related analysis indicated that the area under the ROC curve was 0.8594 (p=0.0157). Conclusion: Here we present, for the first time, the miRNA expression profile of serum exosomes derived from PC patients. Based on this result, we presently suggest that the exosomal hsa-miR-27a-5p may serve as a putative tumor marker for preoperative identification of PC and PA subjects.

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