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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

ABSTRACT: 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.

ORGANISM(S): Camelus bactrianus

PROVIDER: GSE197066 | GEO | 2022/02/23

REPOSITORIES: GEO

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Project description:Purpose：Early diagnosis of nasopharyngeal carcinoma (NPC) is vital to improve the prognosis of these patients. However, early diagnosis of NPC is typically challenging.Therefore, we explored the roles and mechanisms of exosomes in plasma of patients with early-stage NPC. Methods: Exosomes in plasma were extracted by ultra-high-speed centrifugation.Western blot and transmission electron microscopy (TEM) were used to verify the purity of exosomes. The sequencing data (6 plasma samples from healthy volunteers vs. 6 NPC plasma samples) were analyzed by principal component analysis (PCA), DESeq2, gene ontology (GO), kyoto encyclopedia of genes and genomes (KEGG), and TargetScan. The differentially expressed miRNAs (DEmiRNAs) were obtained from the dataset (GSE118720) downloaded from the Gene Expression Omnibus (GEO) repository. Additionally, the datasets downloaded from the GEO database (GSE12452, GSE13597, GSE53819, GSE64634) were used to predict the target genes and functions of hsa-miR-1301-3p. qPCR was applied to verify the differences in the expressions of hsa-miR-1301-3p between 10 normal plasma and 10 NPC plasma samples. Results:Western blot, TEM, and Nanoparticle Tracking Analysis showed adequate purity of the extracted exosomes. RNA-seq analysis revealed 21 upregulated miRNAs, and 10 downregulated miRNAs in plasma exosomes of early-stage NPC patients. GO analysis showed that the target genes of DEmiRNAs were mainly enriched in DNA synthesis and transcription regulation. KEGG analysis revealed that DEmiRNAs were mainly enriched in PI3K-Akt and MAPK signaling pathways. Moreover, the expression of hsa-mir-1301-3p was verified to be significantly upregulated in enlarged samples of plasma exosomes. Conclusions:We identified several DEmiRNAs extracted from tumor-derived exosomes between normal plasma and early-stage NPC plasma. Bioinformatics analyses indicated that these DEmiRNAs may be related to NPC development. Our study may provide novel insights into underlying biomarkers and mechanisms of plasma exosomes in early-stage NPC.

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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

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