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利用RNA测序技术鉴定骨肉瘤发展过程中的差异表达基因。

Identification of differentially expressed genes in the development of osteosarcoma using RNA-seq.

作者信息

Yang Yihao, Zhang Ya, Qu Xin, Xia Junfeng, Li Dongqi, Li Xiaojuan, Wang Yu, He Zewei, Li Su, Zhou Yonghong, Xie Lin, Yang Zuozhang

机构信息

Bone and Soft Tissue Tumors Research Center of Yunnan Province, Department of Orthopaedics, The Third Affiliated Hospital of Kunming Medical University (Tumor Hospital of Yunnan Province), Kunming, Yunnan 650118, China.

出版信息

Oncotarget. 2016 Dec 27;7(52):87194-87205. doi: 10.18632/oncotarget.13554.

Abstract

OBJECTIVE

Osteosarcoma (OS) is a malignant bone tumor with high morbidity in young adults and adolescents. This study aimed to discover potential early diagnosis biomarkers in OS.

RESULTS

In total, 111 differentially expressed genes (DEGs) were identified in primary OS compared with normal controls and 235 DEGs were identified in metastatic OS compared with primary OS. AURKB and PPP2R2B were the significantly up-regulated and down-regulated hub proteins, respectively, in the PPI protein-protein network (PPI) network of primary OS. ISG15 and BTRC were the significantly up-regulated and down-regulated hub proteins, respectively, in the network of metastatic OS. The DEGs in metastatic OS compared with primary OS were significantly enriched in the arachidonic acid metabolism, malaria, and chemokine signaling pathways. Finally, we employed quantitative real-time polymerase chain reaction (qRT-PCR) to validate the expression levels of candidate DEGs and the results indicated that our bioinformatics approach was acceptable.

MATERIALS AND METHODS

The mRNA expression profiling of 20 subjects was obtained through high-throughput RNA-sequencing. DEGs were identified between primary OS and normal Control, and between primary OS and metastatic OS, respectively. Functional annotation and PPI networks were used to obtain insights into the functions of DEGs. qRT-PCR was performed to detect the expression levels of dysregulated genes in OS.

CONCLUSIONS

Our work might provide groundwork for the further exploration of tumorigenesis and metastasis mechanisms of OS.

摘要

目的

骨肉瘤(OS)是一种在青壮年和青少年中发病率较高的恶性骨肿瘤。本研究旨在发现骨肉瘤潜在的早期诊断生物标志物。

结果

与正常对照相比,在原发性骨肉瘤中总共鉴定出111个差异表达基因(DEG);与原发性骨肉瘤相比,在转移性骨肉瘤中鉴定出235个DEG。在原发性骨肉瘤的蛋白质-蛋白质相互作用(PPI)网络中,AURKB和PPP2R2B分别是显著上调和下调的枢纽蛋白。在转移性骨肉瘤网络中,ISG15和BTRC分别是显著上调和下调的枢纽蛋白。与原发性骨肉瘤相比,转移性骨肉瘤中的DEG在花生四烯酸代谢、疟疾和趋化因子信号通路中显著富集。最后,我们采用定量实时聚合酶链反应(qRT-PCR)来验证候选DEG的表达水平,结果表明我们的生物信息学方法是可接受的。

材料与方法

通过高通量RNA测序获得20名受试者的mRNA表达谱。分别在原发性骨肉瘤与正常对照之间以及原发性骨肉瘤与转移性骨肉瘤之间鉴定DEG。使用功能注释和PPI网络来深入了解DEG的功能。进行qRT-PCR以检测骨肉瘤中失调基因的表达水平。

结论

我们的工作可能为进一步探索骨肉瘤的肿瘤发生和转移机制提供基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7c98/5349981/bc8a7f311423/oncotarget-07-87194-g001.jpg

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