Systematic analysis of human oncogenic viruses in colon cancer revealed EBV latency in lymphoid infiltrates
© Fiorina et al.; licensee BioMed Central Ltd. 2014
Received: 30 December 2013
Accepted: 28 April 2014
Published: 3 June 2014
Environmental factors may play a role in colon cancer. In this view, several studies investigated tumor samples for the presence of various viral DNA with conflicting results.
We undertook a systematic DNA analysis of 44 consecutive, prospectively collected primary tumor samples by real time and qualitative PCR for viruses of known or potential oncogenic role in humans, including polyomavirus (JCV, BKV, Merkel cell polyomavirus), HPV, HTLV, HHV-8 and EBV. Negative controls consisted of surgical resection margins. No evidence of genomic DNA fragments from tested virus were detected, except for EBV, which was found in a significant portion of tumors (23/44, 52%). Real-time PCR showed that EBV DNA was present at a highly variable content (median 258 copies in 105 cells, range 15–4837). Presence of EBV DNA had a trend to be associated with high lymphocyte infiltration (p = 0.06, χ 2 test), and in situ hybridization with EBER1-2 probes revealed latency in a fraction of these lymphoid cells, with just a few scattered plasma cells positive for BZLF-1, an immediate early protein expressed during lytic replication. LMP-1 expression was undetectable by immunohistochemistry.
These results argue against a significant involvement of the tested oncogenic viruses in established colon cancer.
It is estimated that 16-18% of the global cancer burden can be associated with oncogenic viruses [1, 2]. The DNA viruses of recognized pathogenic role in humans include HCV, HBV, HPV, HHV-8, MCPyV and EBV, while the role of polyomaviruses JCV and BKV is controversial [1, 3]. Colon cancer is a leading cause of cancer-related death and morbidity in western countries. The pathogenesis of this cancer is highly complex and it involves sequential genetic and epigenetic mechanisms [4, 5]. A possible contribution of environmental agents, including bacteria and viruses, is also considered [6, 7]. However, the search for a pathogenic agent generated conflicting results, possibly related to technical reasons or other unknown factors .
Clinical and pathological features of the colon cancer cases analysed for potential oncogenic viruses
Colon cancer (n = 44)
No. of cases
Median of age = 75 years (range 46–84)
≤ 65 years
≥ 65 years
Sigmoid - Rectal
< 65 years
In conclusion, we performed a PCR-based systematic analysis for potential oncogenic viruses in clinically established colon cancer and EBV was the only one detected. The viral infection was restricted to latency in the lymphoid infiltrate, in line with the few reports that used in situ hybridization with EBER probes [19, 20], while we noted an association with high lymphocyte infiltration, a well-recognized favorable prognostic parameter. EBV positivity in lymphoid infiltrates may occasionally be extensive (Figure 1), much higher than expected on the numbers of circulating EBV positive memory B-lymphocytes in normal individuals, and it might be of interest to studying this phenomenon in specifically designed studies. In summary, the present analysis does not support a significant involvement of the tested viruses in manifest colon cancer, and suggests that new approaches  capable of detecting known and unknown non-human sequences should be investigated to study the role of infectious agents in colon cancer.
The study was supported by Fondazione IRCCS Policlinico “San Matteo”, Ricerca Corrente to VP and FB. ED is recipient of a fellowship from Anatomic Pathology Section, IRCCS Fondazione Policlinico “San Matteo,” Pavia, Italy; RR is a recipient of a grant (project #580) from the Italian Ministry of Health (5 per Mille) of IRCCS Fondazione Policlinico “San Matteo”, Pavia, Italy.
- Bergonzini V, Salata C, Calistri A, Parolin C, Palù G: View and review on viral oncology research. Infect Agent Cancer. 2010, 5: 11-10.1186/1750-9378-5-11.PubMedPubMed CentralView ArticleGoogle Scholar
- de Martel C, Ferlay J, Franceschi S, Vignat J, Bray F, Forman D, Plummer M: Global burden of cancers attributable to infections in 2008: a review and synthetic analysis. Lancet Oncol. 2012, 13 (6): 607-615. 10.1016/S1470-2045(12)70137-7.PubMedView ArticleGoogle Scholar
- Perfetti V, Ricotti M, Buonaguro F, Tirelli U, Pedrazzoli P: An overview of viral oncology in Italy - report from the Pavia meeting on solid tumors. Infect Agent Cancer. 2012, 7 (1): 23-10.1186/1750-9378-7-23.PubMedPubMed CentralView ArticleGoogle Scholar
- Grady WM, Carethers JM: Genomic and epigenetic instability in colorectal cancer pathogenesis. Gastroenterology. 2008, 135 (4): 1079-1099. 10.1053/j.gastro.2008.07.076.PubMedPubMed CentralView ArticleGoogle Scholar
- van Engeland M, Derks S, Smits KM, Meijer GA, Herman JG: Colorectal cancer epigenetics: complex simplicity. J Clin Oncol. 2011, 29 (10): 1382-1391. 10.1200/JCO.2010.28.2319.PubMedView ArticleGoogle Scholar
- Hasan N, Pollack A, Cho I: Infectious causes of colorectal cancer. Infect Dis Clin North Am. 2010, 24 (4): 1019-1039. 10.1016/j.idc.2010.07.009.PubMedView ArticleGoogle Scholar
- Travaglione S, Fabbri A, Fiorentini C: The Rho-activating CNF1 toxin from pathogenic E. coli: a risk factor for human cancer development?. Infect Agent Cancer. 2008, 3: 4-10.1186/1750-9378-3-4.PubMedPubMed CentralView ArticleGoogle Scholar
- Baldanti F, Gatti M, Furione M, Paolucci S, Tinelli C, Comoli P, Merli P, Locatelli F: Kinetics of Epstein-Barr virus DNA load in different blood compartments of pediatric recipients of T-cell-depleted HLA-haploidentical stem cell transplantation. J Clin Microbiol. 2008, 46 (11): 3672-3677. 10.1128/JCM.00913-08.PubMedPubMed CentralView ArticleGoogle Scholar
- Moens B, López G, Adaui V, González E, Kerremans L, Clark D, Verdonck K, Gotuzzo E, Vanham G, Cassar O, Gessain A, Vandamme AM, van Dooren S: Development and validation of a multiplex real-time PCR assay for simultaneous genotyping and human T-lymphotropic virus type 1, 2, and 3 proviral load determination. J Clin Microbiol. 2009, 47 (11): 3682-3691. 10.1128/JCM.00781-09.PubMedPubMed CentralView ArticleGoogle Scholar
- Watzinger F, Suda M, Preuner S, Baumgartinger R, Ebner K, Baskova L, Niesters HG, Lawitschka A, Lion T: Real-time quantitative PCR assays for detection and monitoring of pathogenic human viruses in immunosuppressed pediatric patients. J Clin Microbiol. 2004, 42 (11): 5189-5198. 10.1128/JCM.42.11.5189-5198.2004.PubMedPubMed CentralView ArticleGoogle Scholar
- Saundh BK, Tibble S, Baker R, Sasnauskas K, Harris M, Hale A: Different patterns of BK and JC polyomavirus reactivation following renal transplantation. J Clin Pathol. 2010, 63 (8): 714-718. 10.1136/jcp.2009.074864.PubMedView ArticleGoogle Scholar
- Pancaldi C, Corazzari V, Maniero S, Mazzoni E, Comar M, Martini F, Tognon M: Merkel cell polyomavirus DNA sequences in the buffy coats of healthy blood donors. Blood. 2011, 117 (26): 7099-7101. 10.1182/blood-2010-09-310557.PubMedView ArticleGoogle Scholar
- Qu W, Jiang G, Cruz Y, Chang CJ, Ho GY, Klein RS, Burk RD: PCR detection of human papillomavirus: comparison between MY09/MY11 and GP5+/GP6+ primer systems. J Clin Microbiol. 1997, 35 (6): 1304-1310.PubMedPubMed CentralGoogle Scholar
- Newcomb PA, Bush AC, Stoner GL, Lampe JW, Potter JD, Bigler J: No evidence of an association of JC virus and colon neoplasia. Cancer Epidemiol Biomarkers Prev. 2004, 13 (4): 662-666.PubMedGoogle Scholar
- Militello V, Trevisan M, Squarzon L, Biasolo MA, Rugge M, Militello C, Palù G, Barzon L: Investigation on the presence of polyomavirus, herpesvirus, and papillomavirus sequences in colorectal neoplasms and their association with cancer. Int J Cancer. 2009, 124 (10): 2501-2503. 10.1002/ijc.24224.PubMedView ArticleGoogle Scholar
- Theodoropoulos G, Panoussopoulos D, Papaconstantinou I, Gazouli M, Perdiki M, Bramis J, Lazaris AC: Assessment of JC polyoma virus in colon neoplasms. Dis Colon Rectum. 2005, 48 (1): 86-91. 10.1007/s10350-004-0737-2.PubMedView ArticleGoogle Scholar
- Chiaravalli AM, Longhi E, Vigetti D, De Stefano FI, Deleonibus S, Capella C, Solcia E, Parravicini C: Gastrointestinal cancers reactive for the PAb416 antibody against JCV/SV40 T-Ag lack JCV DNA sequences while showing a distinctive pathologic profile. J Clin Pathol. 2013, 66 (1): 44-49. 10.1136/jclinpath-2012-200963.PubMedView ArticleGoogle Scholar
- Laghi L, Randolph AE, Chauhan DP, Marra G, Major EO, Neel JV, Boland CR: JC virus DNA is present in the mucosa of the human colon and in colorectal cancers. Proc Natl Acad Sci U S A. 1999, 96 (13): 7484-7489. 10.1073/pnas.96.13.7484.PubMedPubMed CentralView ArticleGoogle Scholar
- Liu HX, Ding YQ, Li X, Yao KT: Investigation of Epstein-Barr virus in Chinese colorectal tumors. World J Gastroenterol. 2003, 9 (11): 2464-2468.PubMedPubMed CentralGoogle Scholar
- Wong NA, Herbst H, Herrmann K, Kirchner T, Krajewski AS, Moorghen M, Niedobitek F, Rooney N, Shepherd NA, Niedobitek G: Epstein-Barr virus infection in colorectal neoplasms associated with inflammatory bowel disease: detection of the virus in lymphomas but not in adenocarcinomas. J Pathol. 2003, 201 (2): 312-318. 10.1002/path.1442.PubMedView ArticleGoogle Scholar
- Barzon L, Lavezzo E, Costanzi G, Franchin E, Toppo S, Palù G: Next-generation sequencing technologies in diagnostic virology. J Clin Virol. 2013, 58 (2): 346-350. 10.1016/j.jcv.2013.03.003.PubMedView ArticleGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.