Microrna Profiling of Serum Exosomes in Patients with Osteosarcoma by High-Throughput Sequencing

骨肉瘤 微泡 小RNA 外体 流式细胞术 下调和上调 免疫印迹 癌症研究 体内 生物 分子生物学 基因 生物化学 生物技术
作者
Zhimeng Ye,Zhuojun Zheng,Linrui Peng
出处
期刊:Journal of Investigative Medicine [SAGE Publishing]
卷期号:68 (4): 893-901 被引量:23
标识
DOI:10.1136/jim-2019-001196
摘要

The microRNA expression profile of plasma exosomes in osteosarcoma needs to be further explored. The present study intends to investigate the practicality of plasma exosomal miRNAs as novel biomarkers of osteosarcoma. In the study, exosome-like vesicles were purified from the plasma of patients with osteosarcoma and healthy control. Differential centrifugation was used. The purified vesicles which ranged from 50 to 100 nm in size were identified as exosomes by transmission electron microscopy and western blot. Validating assays in vitro and in vivo were performed via CCK8, reverse transcription-quantitative PCR, flow cytometry, transwell and wound healing assays and xenograft model. High-throughput sequencing identified that 57 miRNAs, 20 of which were upregulated and 37 downregulated, were differentially expressed in patients with osteosarcoma and healthy control (p<0.01; fold change ≥3). In comparison to the controls, the expression levels of miR-92a-3p, miR-130a-3p, miR-195-3 p, miR-335-5 p, let-7i-3p were upregulated in the exosomes from patients with osteosarcoma with statistical significance. Studies in vitro and in vivo have proved that osteosarcoma-secreted exosomes from miR-195-3 p upregulated 143B osteosarcoma cells promote cell proliferation and invasion. Overall, the present study identified exosomal miRNAs with dysregulated expression in patients with osteosarcoma, and they may have potential as targets for the treatment of patients with osteosarcoma.

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