Identification of Altered Circular RNA Expression in Serum Exosomes from Patients with Papillary Thyroid Carcinoma by High-Throughput Sequencing

利用高通量测序技术鉴定乳头状甲状腺癌患者血清外泌体中环状RNA表达的改变

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Abstract

BACKGROUND This study aimed to identify altered exosome circular RNA (circRNA) in the serum of patients with papillary thyroid carcinoma using high-throughput sequencing. MATERIAL AND METHODS Serum was collected from three patients with papillary thyroid carcinoma and three patients with a benign thyroid goiter. Exosomes were isolated using an exosome isolation kit and confirmed by transmission electron microscopy. Exosome circRNAs were analyzed by high-throughput sequencing using the HiSeq 4000 sequencer. The differentially expressed circRNAs were confirmed by fluorescence quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS Twenty-two differentially expressed circRNAs were screened, which included three that were upregulated and 19 that were down-regulated in serum from patients with papillary thyroid carcinoma compared with controls. Gene Ontology (GO) enrichment analysis showed that these differentially expressed circRNAs were associated with 16 signaling pathways, including the thyroid hormone signaling pathway, the PI3K-Akt signaling pathway, and the AMPK signaling pathway. Three differentially regulated circRNAs included hsacirc_007293, hsacirc_031752, and hsacirc_020135 were confirmed by qRT- PCR. The expression trends were consistent between the high-throughput sequencing technique and qRT-PCR. CONCLUSIONS The findings from this study have shown that gene regulation can be studied from exosomes obtained from serum samples in patients with papillary thyroid carcinoma, and supports the need for further studies on the role of exosome circRNAs in thyroid cancer.

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