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Journal of Virology, March 2009, p. 2357-2367, Vol. 83, No. 5
0022-538X/09/$08.00+0     doi:10.1128/JVI.02104-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Comprehensive Profiling of Epstein-Barr Virus MicroRNAs in Nasopharyngeal Carcinoma{triangledown} ,{dagger}

Katherine Cosmopoulos,1 Michiel Pegtel,2 Jared Hawkins,1 Howell Moffett,3,4 Carl Novina,3,4 Jaap Middeldorp,2 and David A. Thorley-Lawson1*

Department of Pathology, Tufts University School of Medicine, Jaharis Building, Boston, Massachusetts,1 Department of Pathology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands,2 Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, Massachusetts,3 Department of Pathology, Harvard Medical School, Boston, Massachusetts4

Received 6 October 2008/ Accepted 11 December 2008

Epstein-Barr Virus (EBV) establishes a long-term latent infection and is associated with a number of human malignancies that are thought to arise from deregulation of different stages of the viral life cycle. Recently, a large number of microRNAs (miRNAs) have been described for EBV, and it has been suggested that their expression may vary between the different latency states found in normal and malignant tissue. To date, however, no technique has been utilized to comprehensively and quantitatively test this idea by profiling expression of the EBV miRNAs in primary infected tissues. We describe here a multiplex reverse transcription-PCR assay that allows the profiling of 39 of the 40 known mature EBV miRNAs from as little as 250 ng of RNA. With this approach, we present a comprehensive profile of EBV miRNAs in primary nasopharyngeal carcinoma (NPC) tumors including estimates of miRNA copy number per tumor cell. This is the first comprehensive profiling of EBV miRNAs in any EBV-associated tumor. In contrast to previous suggestions, we show that the BART-derived miRNAs are present in a wide range of copy numbers from ≤103 per cell in both primary tumors and the widely used NPC-derived C666-1 cell line. However, we confirm the hypothesis that the BHRF1 miRNAs are not expressed in NPC. Lastly, we demonstrate that EBV miRNA expression in the widely used NPC line C666-1 is, with some caveats, broadly representative of primary NPC tumors.

* Corresponding author. Mailing address: Department of Pathology, Jaharis Building, Tufts University School of Medicine, 150 Harrison Ave., Boston, MA 02111. Phone: (617) 636-2726. Fax: (617) 636-2990. E-mail: david.thorley-lawson{at}tufts.edu

{triangledown} Published ahead of print on 17 December 2008.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, March 2009, p. 2357-2367, Vol. 83, No. 5
0022-538X/09/$08.00+0     doi:10.1128/JVI.02104-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Zhu, J. Y., Pfuhl, T., Motsch, N., Barth, S., Nicholls, J., Grasser, F., Meister, G. (2009). Identification of Novel Epstein-Barr Virus MicroRNA Genes from Nasopharyngeal Carcinomas. J. Virol. 83: 3333-3341 [Abstract] [Full Text]  


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