Abstract
BackgroundEarly 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 pathogenetic roles and associated mechanisms of exosomes in plasma of patients with early-stage NPC.MethodsExosomes 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.ResultsWestern 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.ConclusionsWe 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.
Highlights
Diagnosis of nasopharyngeal carcinoma (NPC) is vital to improve the prognosis of these patients
Isolation and characterization of exosomes Exosomes are microvesicles that can transport miRNAs, mRNAs and proteins, which are essential for the cell to cell communication
These results indicated the high efficiency of ultra-high-speed centrifugation in extracting exosomes
Summary
Diagnosis of nasopharyngeal carcinoma (NPC) is vital to improve the prognosis of these patients. Early diagnosis of NPC is typically challenging. We explored the pathogenetic roles and associated mechanisms of exosomes in plasma of patients with early-stage NPC. Nasopharyngeal carcinoma (NPC) is a malignant cancer that shows wide variability in geographical distribution. More than 70% of NPC patients are diagnosed at an advanced stage [5, 6]. Early diagnosis of NPC is vital to improve the prognosis of these patients. Nasopharyngeal biopsy and imaging examination are the main modalities for the diagnosis of NPC. Patients with early-stage NPC are typically asymptomatic and are unlikely to be subjected to these investigations; biopsy is an invasive procedure. Unraveling the mechanisms of the occurrence and development of early-stage NPC and identification of potential molecular biomarkers for early diagnosis of NPC are key research imperatives
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