Alterations in cellular expression in EBV infected epithelial cell lines and tumors

Autoři: Rachel Hood Edwards aff001;  Robert Dekroon aff001;  Nancy Raab-Traub aff001
Působiště autorů: Lineberger Comprehensive Cancer Center University of North Carolina at Chapel Hill Chapel Hill, North Carolina, United States of America aff001;  Department of Microbiology & Immunology University of North Carolina at Chapel Hill Chapel Hill, North Carolina, United States of America aff002
Vyšlo v časopise: Alterations in cellular expression in EBV infected epithelial cell lines and tumors. PLoS Pathog 15(10): e1008071. doi:10.1371/journal.ppat.1008071
Kategorie: Research Article


The Epstein Barr virus (EBV) is linked to the development of two major epithelial malignancies, gastric carcinoma and nasopharyngeal carcinoma. This study evaluates the effects of EBV on cellular expression in a gastric epithelial cell line infected with or without EBV and a nasopharyngeal carcinoma cell line containing EBV. The cells were grown in vitro and as tumors in vivo. The effects on cellular expression were determined using both 2D DIGE proteomics and high throughput RNA sequencing. The data identify multiple pathways that were uniquely activated in vitro. RNA sequences mapping to the mouse genome were identified in both the EBV positive and negative tumor samples in vivo, although, differences between the EBV positive and negative cells were not apparent. However, the tumors appeared to be grossly distinct. The majority of the identified canonical pathways based on two fold changes in expression had decreased activity within the tumors in vivo. Identification of the predicted upstream regulating factors revealed that in vitro the regulating factors were primarily protein transcriptional regulators. In contrast, in vivo the predicted regulators were frequently noncoding RNAs. Hierarchical clustering distinguished the cell lines and tumors, the EBV positive tumors from the EBV negative tumors, and the NPC tumors from the gastric tumors and cell lines. The delineating genes were changed greater than 4 fold and were frequently regulated by protein transcription factors. These data suggest that EBV distinctly affects cellular expression in gastric tumors and NPC and that growth in vivo requires activation of fewer cellular signaling pathways. It is likely that the broad changes in cellular expression that occur at low levels are controlled by regulatory viral and cellular RNAs while major changes are affected by induced protein regulators.

Klíčová slova:

Enzyme regulation – Epstein-Barr virus – Gene expression – Gene regulation – MicroRNAs – Regulator genes – Transcriptional control – Nasopharyngeal carcinoma


1. Raab-Traub N. EBV-induced oncogenesis. In: Arvin A, Campadelli-Fiume G, Mocarski E, Moore PS, Roizman B, Whitley R, et al., editors. Human Herpesviruses: Biology, Therapy, and Immunoprophylaxis. Cambridge2007.

2. Busson P, Ganem G, Flores P, Mugneret F, Clausse B, Caillou B, et al. Establishment and characterization of three transplantable EBV-containing nasopharyngeal carcinomas. International journal of cancer Journal international du cancer. 1988;42(4):599–606. doi: 10.1002/ijc.2910420422 2971626

3. Yip YL, Lin W, Deng W, Jia L, Lo KW, Busson P, et al. Establishment of a nasopharyngeal carcinoma cell line capable of undergoing lytic Epstein-Barr virus reactivation. Laboratory investigation; a journal of technical methods and pathology. 2018;98(8):1093–104. doi: 10.1038/s41374-018-0034-7 29769697

4. Cheung ST, Huang DP, Hui AB, Lo KW, Ko CW, Tsang YS, et al. Nasopharyngeal carcinoma cell line (C666-1) consistently harbouring Epstein-Barr virus. International journal of cancer Journal international du cancer. 1999;83(1):121–6. doi: 10.1002/(sici)1097-0215(19990924)83:1<121::aid-ijc21>;2-f 10449618

5. Huang J, Liao G, Chen H, Wu FY, Hutt-Fletcher L, Hayward GS, et al. Contribution of C/EBP proteins to Epstein-Barr virus lytic gene expression and replication in epithelial cells. Journal of virology. 2006;80(3):1098–109. doi: 10.1128/JVI.80.3.1098-1109.2006 16414987

6. Raab-Traub N. Nasopharyngeal Carcinoma: An Evolving Role for the Epstein-Barr Virus. Current topics in microbiology and immunology. 2015;390:339–63. doi: 10.1007/978-3-319-22822-8_14 26424653

7. Kieff E, and Rickinson A. B. Epstein-Barr Virus and Its Replication. In: Howley DMKaPM, editor. Field's Virology. II. Fourth ed. Philadelphia, PA: Lippincott Williams & Wilkins Publishers; 2001. p. 2511–73.

8. Marquitz AR, Mathur A, Shair KH, Raab-Traub N. Infection of Epstein-Barr virus in a gastric carcinoma cell line induces anchorage independence and global changes in gene expression. Proceedings of the National Academy of Sciences of the United States of America. 2012;109(24):9593–8. doi: 10.1073/pnas.1202910109 22647604

9. Marquitz AR, Mathur A, Chugh PE, Dittmer DP, Raab-Traub N. Expression profile of microRNAs in Epstein-Barr virus-infected AGS gastric carcinoma cells. Journal of virology. 2014;88(2):1389–93. doi: 10.1128/JVI.02662-13 24227849

10. Cai X, Schafer A, Lu S, Bilello JP, Desrosiers RC, Edwards R, et al. Epstein-Barr virus microRNAs are evolutionarily conserved and differentially expressed. PLoS pathogens. 2006;2(3):e23. doi: 10.1371/journal.ppat.0020023 16557291

11. Marquitz AR, Mathur A, Edwards RH, Raab-Traub N. Host Gene Expression Is Regulated by Two Types of Noncoding RNAs Transcribed from the Epstein-Barr Virus BamHI A Rightward Transcript Region. Journal of virology. 2015;89(22):11256–68. doi: 10.1128/JVI.01492-15 26311882

12. Batista PJ, Chang HY. Long noncoding RNAs: cellular address codes in development and disease. Cell. 2013;152(6):1298–307. doi: 10.1016/j.cell.2013.02.012 23498938

13. Qiu J, Smith P, Leahy L, Thorley-Lawson DA. The Epstein-Barr Virus Encoded BART miRNAs Potentiate Tumor Growth In Vivo. PLoS pathogens. 2015;11(1):e1004561. doi: 10.1371/journal.ppat.1004561 25590614

14. Barranco SC, Townsend CM Jr., Casartelli C, Macik BG, Burger NL, Boerwinkle WR, et al. Establishment and characterization of an in vitro model system for human adenocarcinoma of the stomach. Cancer research. 1983;43(4):1703–9. 6831414

15. Burkovics P, Hajdu I, Szukacsov V, Unk I, Haracska L. Role of PCNA-dependent stimulation of 3'-phosphodiesterase and 3'-5' exonuclease activities of human Ape2 in repair of oxidative DNA damage. Nucleic acids research. 2009;37(13):4247–55. doi: 10.1093/nar/gkp357 19443450

16. Ungerleider NA, Jain V, Wang Y, Maness NJ, Blair RV, Alvarez X, et al. Comparative Analysis of Gammaherpesvirus Circular RNA Repertoires: Conserved and Unique Viral Circular RNAs. Journal of virology. 2019;93(6).

17. Henderson S, Huen D, Rowe M, Dawson C, Johnson G, Rickinson A. Epstein-Barr virus-coded BHRF1 protein, a viral homologue of Bcl-2, protects human B cells from programmed cell death. Proceedings of the National Academy of Sciences of the United States of America. 1993;90(18):8479–83. doi: 10.1073/pnas.90.18.8479 8397406

18. Vereide D, Sugden B. Proof for EBV's sustaining role in Burkitt's lymphomas. Seminars in cancer biology. 2009;19(6):389–93. doi: 10.1016/j.semcancer.2009.07.006 19628040

19. Xiang T, Lin YX, Ma W, Zhang HJ, Chen KM, He GP, et al. Vasculogenic mimicry formation in EBV-associated epithelial malignancies. Nat Commun. 2018;9(1):5009. doi: 10.1038/s41467-018-07308-5 30479336

20. Scholle F, Bendt KM, Raab-Traub N. Epstein-Barr virus LMP2A transforms epithelial cells, inhibits cell differentiation, and activates Akt. Journal of virology. 2000;74(22):10681–9. doi: 10.1128/jvi.74.22.10681-10689.2000 11044112

21. Skalsky RL, Corcoran DL, Gottwein E, Frank CL, Kang D, Hafner M, et al. The viral and cellular microRNA targetome in lymphoblastoid cell lines. PLoS pathogens. 2012;8(1):e1002484. doi: 10.1371/journal.ppat.1002484 22291592

22. Marquitz AR, Raab-Traub N. The role of miRNAs and EBV BARTs in NPC. Seminars in cancer biology. 2012;22(2):166–72. doi: 10.1016/j.semcancer.2011.12.001 22178394

23. [A new nickel-chromium: Phoenix Superalloy]. Les Cahiers de prothese. 1975(11):143–4. 1072859

24. Mainou BA, Raab-Traub N. LMP1 strain variants: biological and molecular properties. Journal of virology. 2006;80(13):6458–68. doi: 10.1128/JVI.00135-06 16775333

25. Afgan E, Baker D, van den Beek M, Blankenberg D, Bouvier D, Cech M, et al. The Galaxy platform for accessible, reproducible and collaborative biomedical analyses: 2016 update. Nucleic acids research. 2016;44(W1):W3–W10. doi: 10.1093/nar/gkw343 27137889

26. Torre D, Lachmann A, Ma'ayan A. BioJupies: Automated Generation of Interactive Notebooks for RNA-Seq Data Analysis in the Cloud. Cell Syst. 2018;7(5):556–61 e3. doi: 10.1016/j.cels.2018.10.007 30447998

27. Mainou BA, Everly DN, Jr., Raab-Traub N. Epstein-Barr virus latent membrane protein 1 CTAR1 mediates rodent and human fibroblast transformation through activation of PI3K. Oncogene. 2005;24(46):6917–24. doi: 10.1038/sj.onc.1208846 16007144

28. Marquitz AR, Mathur A, Nam CS, Raab-Traub N. The Epstein-Barr Virus BART microRNAs target the pro-apoptotic protein Bim. Virology. 2011;412(2):392–400. doi: 10.1016/j.virol.2011.01.028 21333317

29. Ünlü M, Morgan ME, & Minden JS. Difference gel electrophoresis. A single gel method for detecting changes in protein extracts. Electrophoresis. 1997;18(11):2071–2077. doi: 10.1002/elps.1150181133 9420172

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