Comprehensive profiling of EBV gene expression in nasopharyngeal carcinoma through paired-end transcriptome sequencing

Abstract

The latent expression pattern of Epstein-Barr Virus (EBV) genes in nasopharyngeal carcinoma (NPC) has been extensively investigated, and the expression of several lytic genes in NPC has been reported. However, comprehensive information through EBV transcriptome analysis in NPC is limited. We performed paired-end RNA-seq to systematically and comprehensively characterize the expression of EBV genes in NPC tissue and C666-1 NPC cell line, which consistently carries EBV. In addition to the transcripts restricted to type II latency infection, the type III latency EBNA3s genes and a substantial number of lytic genes, such as BZLF1, BRLF1, and BMRF1, were detected through RNA-seq and were further verified in C666-1 cells and NPC tissue through real-time PCR. We also performed clustering analysis to classify NPC patient groups in terms of EBV gene expression, which presented two subtypes of NPC samples. Results revealed interesting patterns of EBV gene expression in NPC patients. This clustering was correlated with many signaling pathways, such as those related to heterotrimeric G-protein signaling, inflammation mediated by chemokine and cytokine signaling, ribosomes, protein metabolism, influenza infection, and ECM-receptor interaction. Our combined findings suggested that the expression of EBV genes in NPC is restricted not only to type II latency genes but also to type III latency and lytic genes. This study provided further insights into the potential role of EBV in the development of NPC.

Keywords

Epstein-Barr virus / paired-end transcriptome sequencing / latency genes / lytic genes / nasopharyngeal carcinoma

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Acknowledgements

This study was supported by 973 Program and 863 Program from the Ministry of Science and Technology of China (Nos. 2011CB504300 and 2015AA020931), National Natural Science Foundation of China (Nos. 91440106, 91019015, and 81302037), Ph.D. Startup of Guangzhou Medical University (No. 2012C65), Guangzhou Science and Technology Planing Project (No. 2014J4100181), and Young Teacher Training Program of Sun Yat-sen University (No. 13ykpy50).

Compliance with ethics guidelines

Lijuan Hu, Zhirui Lin, Yanheng Wu, Juqin Dong, Bo Zhao, Yanbing Cheng, Peiyu Huang, Lihua Xu, Tianliang Xia, Dan Xiong, Hongbo Wang, Manzhi Li, Ling Guo, Elliott Kieff, Yixin Zeng, Qian Zhong, and Musheng Zeng declare that they have no conflict of interest. All procedures were in accordance with the ethical standards of the respective committees on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000 (5). Informed consent was obtained from all patients who were included in the study.

Electronic Supplementary Material ƒSupplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s11684-016-0436-0 and is accessible for authorized users.

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