Comprehensive Profiling of Epstein-Barr Virus MicroRNAs in Nasopharyngeal Carcinoma

被引:141
作者
Cosmopoulos, Katherine [1 ]
Pegtel, Michiel [2 ]
Hawkins, Jared [1 ]
Moffett, Howell [3 ,4 ]
Novina, Carl [3 ,4 ]
Middeldorp, Jaap [2 ]
Thorley-Lawson, David A. [1 ]
机构
[1] Tufts Univ, Sch Med, Dept Pathol, Boston, MA 02111 USA
[2] Vrije Univ Amsterdam, Med Ctr, Dept Pathol, Amsterdam, Netherlands
[3] Dana Farber Canc Inst, Boston, MA 02115 USA
[4] Harvard Univ, Sch Med, Dept Pathol, Boston, MA 02115 USA
关键词
BAMHI-A REGION; PROTEIN EXPRESSION; EBV; TRANSCRIPTION; IDENTIFICATION; INFECTION; FRAGMENT; RNAS; LMP1;
D O I
10.1128/JVI.02104-08
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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 <= 10(3) 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.
引用
收藏
页码:2357 / 2367
页数:11
相关论文
共 40 条
[1]  
Ambros Victor, 2004, Methods Mol Biol, V265, P131
[2]   2 FAMILIES OF SEQUENCES IN THE SMALL RNA-ENCODING REGION OF EPSTEIN-BARR VIRUS (EBV) CORRELATE WITH EBV TYPE-A AND TYPE-B [J].
ARRAND, JR ;
YOUNG, LS ;
TUGWOOD, JD .
JOURNAL OF VIROLOGY, 1989, 63 (02) :983-986
[3]   Epstein-Barr virus-encoded microRNA miR-BART2 down-regulates the viral DNA polymerase BALF5 [J].
Barth, Stephanie ;
Pfuhl, Thorsten ;
Mamiani, Alfredo ;
Ehses, Claudia ;
Roemer, Klaus ;
Kremmer, Elisabeth ;
Jaeker, Christoph ;
Hoeck, Julia ;
Meister, Gunter ;
Graesser, Friedrich A. .
NUCLEIC ACIDS RESEARCH, 2008, 36 (02) :666-675
[4]   TRANSCRIPTS FROM THE EPSTEIN-BARR-VIRUS BAMHI A FRAGMENT ARE DETECTABLE IN ALL 3 FORMS OF VIRUS LATENCY [J].
BROOKS, LA ;
LEAR, AL ;
YOUNG, LS ;
RICKINSON, AB .
JOURNAL OF VIROLOGY, 1993, 67 (06) :3182-3190
[5]   Epstein-Barr virus MicroRNAs are evolutionarily conserved and differentially expressed [J].
Cai, Xuezhong ;
Schafer, Alexandra ;
Lu, Shihua ;
Bilello, John P. ;
Desrosiers, Ronald C. ;
Edwards, Rachel ;
Raab-Traub, Nancy ;
Cullen, Bryan R. .
PLOS PATHOGENS, 2006, 2 (03) :236-247
[6]   Human microRNAs are processed from capped, polyadenylated transcripts that can also function as mRNAs [J].
Cai, XZ ;
Hagedorn, CH ;
Cullen, BR .
RNA, 2004, 10 (12) :1957-1966
[7]   Epstein-Barr virus latent membrane protein 1 induces cellular MicroRNA miR-146a, a modulator of lymphocyte signaling pathways [J].
Cameron, Jennifer E. ;
Yin, Qinyan ;
Fewell, Claire ;
Lacey, Michelle ;
McBride, Jane ;
Wang, Xia ;
Lin, Zhen ;
Schaefer, Brian C. ;
Flemington, Erik K. .
JOURNAL OF VIROLOGY, 2008, 82 (04) :1946-1958
[8]   Real-time quantification of microRNAs by stem-loop RT-PCR [J].
Chen, CF ;
Ridzon, DA ;
Broomer, AJ ;
Zhou, ZH ;
Lee, DH ;
Nguyen, JT ;
Barbisin, M ;
Xu, NL ;
Mahuvakar, VR ;
Andersen, MR ;
Lao, KQ ;
Livak, KJ ;
Guegler, KJ .
NUCLEIC ACIDS RESEARCH, 2005, 33 (20) :e179.1-e179.9
[9]   TRANSCRIPTION OF BAMHI-A REGION OF THE EBV-GENOME IN NPC TISSUES AND B-CELLS [J].
CHEN, HL ;
LUNG, MML ;
SHAM, JST ;
CHOY, DTK ;
GRIFFIN, BE ;
NG, MH .
VIROLOGY, 1992, 191 (01) :193-201
[10]   Epstein-Barr virus BART microRNAs are produced from a large intron prior to splicing [J].
Edwards, Rachel Hood ;
Marquitz, Aron R. ;
Raab-Traub, Nancy .
JOURNAL OF VIROLOGY, 2008, 82 (18) :9094-9106