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CircRNA-sequencing analysis of plasma exosomes in hepatocellular carcinoma

ABSTRACT: circRNA-sequencing analysis of plasma exosomes in hepatocellular carcinoma

PROVIDER: PRJNA692609 | ENA |

REPOSITORIES: ENA

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circRNA-sequencing analysis of plasma exosomes in hepatocellular carcinoma

Project description:Hepatocellular carcinoma (HCC) is currently the third leading cause of death worldwide and the most common type of primary liver cancer, finding noninvasive biomarkers for HCC diagnostic and prognostic are very urging. Previous genomic studies mainly focus on finding miRNA biomarkers for HCC. In this study, we focus on finding circRNA fragments suitable for serving as hcc biomarkers with plasma exosomal RNA-seq..

2021-12-01 | GSE164953 | GEO

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Project description:Circular RNAs (circRNAs) are recently found to be promising kind of biomarkers for tumors. However, plasma exosomal circRNAs in the diagnosis of hepatocellular carcinoma (HCC) is largely unknown. In this study, we report the application of next-generation sequencing technology for high-throughput profiling of plasma exosome general transcriptome between 3 cases of hepatocellular carcinoma patients and 3 cases of healthy control. We obtained exosomes through ultrahigh-speed centrifugation techniques and verified exosomes through immunoblotting, NanoSight NS300(Malvern Panalytical, Malvern, UK) and JEM-1230 transmission electron microscope (TEM) (JEOL, Tokyo, Japan). And then, we find that there are 41 circRNAs that have differential expression. This study discovered the differences between hepatocellular carcinoma and healthy people preliminarily.

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circRNA expression profiles for 10 human samples derived from patients with hepatocellular carcinoma

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Effects of plasma-derived exosomes from the normal and thin camels on hepatocellular carcinoma and their differences at transcriptome and proteomics levels

Project description:Background: Hepatocellular carcinoma (HCC) is a common malignant primary tumor. Camels have high economic and social values, but their potential medical value has not been studied. This study aimed to investigate the effects of thin and normal camel plasma-derived exosomes on HCC. Methods: Plasma was obtained from thin and normal camels, and used to isolate exosomes by ultracentrifugation. The exosomes were then characterized by transmission electron microscopy and Nano particle tracking analyzer. In vivo imaging of nude mice and hematoxylin eosin (HE) staining of liver tissues were used to explore the effects of the exosomes on tumor growth. Finally, the differences of the two exosomes were further analyzed using small RNA sequencing and proteomics. Results: In vivo imaging and HE staining showed that no significant differences were found in fluorescence value and liver tissue morphology between the control mice and the mice treated with the exosomes from thin camels; while the fluorescence value and the live histology changes were alleviated in the mice with the exosomes from normal camels. After sequencing and proteomic analysis, 40 DE-miRNAs (15 down-regulated and 25 up-regulated) and 172 DEPs (77 up-regulated and 95 down-regulated) were identified in the plasma-derived exosomes from normal camels. These identified DE-miRNAs and DEPs were significantly enriched in many signaling pathways. Conclusions: Normal camel plasma-derived exosomes may inhibit the growth of HCC cells through regulating pathways of Ras, Rap1, PI3K-Akt, MAPK, AMPK, and canonical Wnt signaling pathways.

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Exosomal circRNA-MANBA mediates sorafenib resistance of hepatocellular carcinoma via miR-1290/CD109/p-STAT3 (Ser727) axis

Project description:Circular RNAs (circRNAs) are increasingly gaining importance and attention due to their diverse potential functions. Our study aims to explore the novel mechanisms by which exosome-contained circRNAs promote sorafenib resistance in hepatocellular carcinoma (HCC). Here we report that a circular RNA, circRNA-MANBA (a circular RNA upregulated in exosomes derived from sorafenib-resistant HCC cells), plays a significant role in sorafenib resistance in HCC. We identified that circRNA-MANBA is increased in sorafenib-resistant HCC cells, their exosomes, and tumor samples from sorafenib-treated HCC patients. Depletion of circRNA-MANBA substantially increases the cell-killing ability of sorafenib. Co-culture experiments revealed that exosomes from sorafenib-resistant HCC cells carrying large amounts of circRNA-MANBA could transmit drug resistant capacity to parental cells. Further studies revealed that circRNA-MANBA acted as ‘miRNAs sponge’ to absorb miR-1290, which prevented miR-1290 interaction with CD109 and thus upregulated STAT3 phosphorylation subsequently. Using different HCC mouse models, we demonstrated that silencing circRNA-MANBA by injection of siRNA could substantially overcome sorafenib resistance. Our study provides a proof-of-concept demonstration for a potential strategy to overcome sorafenib resistance in HCC patients by targeting circRNA-MANBA.

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RNA-seq reveals abundant circRNA, lncRNA and mRNA in blood exosomes of patients with hepatocellular carcinoma

Project description:Exosomes are small membrane vesicles of endocytic origin secreted by most cells, and contain a wealthy cargo of protein and RNA species that can modulate recipient cells’ behaviors and may be used as biomarkers for diagnosis of human diseases. They have been found in blood and are valuable sources for biomarkers due to selective cargo loading and resemblance to their parental cells. The goal of this study is to identify circRNA, lncRNA and mRNA profiles in human blood by high-throughput RNA sequencing (RNA-seq). 1-4 ml plasma or serum were used to extract exosomal RNAs by exoRNeasy Serum/Plasma Maxi kit (Qiagen). The exosomal RNAs were further treated with DNAse I and subjected to ribosome minus low-input RNAseq library preparation. The libraries were sequenced by Illumina Hiseq platform.

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