Analysis of Epstein Barr Virus Encoded RNA Expression in Nasopharyngeal Carcinoma in North-Eastern India: A Chromogenic in Situ Hybridization Based Study

Document Type : Original


1 Department of Pathology, Post Graduate Institute of Medical Education and Research, Chandigarh, India.

2 Department of Pathology, North Eastern Indira Gandhi Regional Institute of Health and Medical Sciences, Shillong, India.

3 Department of Otorhinolaryngology, North Eastern Indira Gandhi Regional Institute of Health and Medical Sciences, Shillong, India.


Nasopharyngeal carcinoma (NPC) is a common cancer in the North-East region of India. Though the role of environmental contributors of NPC in the North-Eastern part of India is firmly established, EBV as an etiological agent in the region remains unexplored.
Material and Methods:
Fifty-one patients, who presented at the department of ENT, NEIGRIHMS and were confirmed as NPC upon histopathological examination, were included in the study. Chromogenic in-situ hybridization (CISH) was used for the evaluation of EBER (Epstein Barr Virus Encoded RNA). Presence of nuclear signals was taken as positive for EBER expression. EBER status was correlated with various clinicopathological parameters like age, sex, dietary habits, histological types of NPC, and ethnicity of the patients.
The age range of the study group was 25 to 70 years with a mean age of 44.64 years and a male:female ratio of 3:2. Non-keratinizing undifferentiated type of NPC was the most common histological type. EBV was positive in 59% (30/51) of our cases. It showed a statistically significant correlation with the Naga community (P=0.01), with consumption of smoked food (P=0.02), and cigarette smoking (P=0.02). There was no correlation of EBV with age, sex, lymph node metastasis, stage, and histology.
Our result indicates that EBV may be an additional risk factor in the pathogenesis of NPC in this region of India. So apart from lifestyle modification, a future study for a screening test for EBV viral load even in asymptomatic patients may be considered, for determination of disease susceptibility, early diagnosis, and proper management.


Main Subjects

1. Sharma TD, Singh TT, Laishram RS, Sharma LDC, Sunita AK, Imchen LT. Nasopharyngeal Carcinoma- A Clinico-pathological Study in a Regional Cancer Centre of  North-eastern  India. Asian Pacific J Cancer. 2011; 12: 1583-7.
2. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011; 61: 69-90.
3. Ferley J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, method and major patterns in GLOBOCAN 2012. Int J Cancer. 2015; 136(5): 359-86.
4. Kumar S , Zinyu R , Singh IKK , Medhi SB, Baruah T, Das B, et al. Studies on Nasopharyngeal cancer with reference to the North-Eastern region of India. Ann Natl Acad Med Sci. 1996; 32:199.
5. Wei KR, Xu Y, Liu J, Zhang WJ, Liang ZH. Histopathological Classification of Nasopharyngeal Carcinoma. Asian Pacific J Cancer. 2011; 12:
6. Krishna SM , James S, Sreelekha TT, Kattoor J,  Balaram P. Primary Nasopharyngeal Cancer of Indian Origin and the Viral Link - A Preliminary Study. Asian J of Cancer. 2006; 5: 241-52.
7. Liu MT, Hsieh CY, Chang TH, Lin JP, Huang CC, Wang AY. Prognostic factor affecting the outcome of nasopharyngeal carcinoma. Jpn J clin oncol. 2003; 33(10): 501-8.
8. Jitani AK , Raphael V , Mishra J , Shunyu NB, Khonglah Y, Medhi J . Analysis of Human Papilloma Virus 16/18 DNA and its correlation with p16 expression in oral cavity squamous cell carcinoma in North Eastern India : A chromogenic in situ hybridization based study. J Clin Diagn Res. 2015; 9(8): EC04-7.
9. Chang ET, Adami HO. The Enigmatic Epidemiology of Nasopharyngeal Carcinoma. Cancer Epidemiol Biomarkers. 2006; 15: 1765-77.
10. Macmohan B. Epidemiology and Hodgkin disease. Cancer Res. 1966; 26: 1189-201.
11. Brooks L, Yao QY, Rickinson AB, Young LS. Epstein Barr Virus latent gene transcription in nasopharyngeal carcinoma cells: co-expression of EBNA1, LMP1 and LMP2 transcripts. J Virol. 1992; 66(5): 2689-97.
12. Pathmanathan R, Prasad U, Chandrika G, Sadler R, Flynn K, Raab-Traub N. Undifferentiated, nonkeratinizing, and squamous cell carcinoma of the nasopharynx. Am J Pathol. 1995; 146(6): 1355-67.
13. Cheng YJ, Hildesheim A, Hsu MM, Chen IH,  A. Brinton L, H. Levine O, et al. Cigarette smoking, alcohol consumption and risk of nasopharyngeal carcinoma in Taiwan. Cancer Causes Control. 1999; 10(3): 201-7.
14. Chen L, Gallicchio L, Boyd-Lindsley  K, Tao XG, Robinson KA, Lam TK, et al. Alcohol consumption and the risk of Nasopharyngeal carcinoma: A systemic review. Nutr Cancer. 2008; 61: 1-15.
15. Kataki ACSimons MJDas AKSharma KMehra NK. Nasopharyngeal carcinoma in the North-eastern states of India. Chin J Cancer. 2011; 30(2): 106-13.
16. Tsai ST, Jin YT, Mann RB, Ambinder RF. Epstein-barr virus detection in nasopharyngeal tissues of patients with suspected nasopharyngeal carcinoma. Cancer. 1998; 82(8): 1449-53.
17. Wu TC, Mann RB, Epstein IJ,  MacMahon E, Lee WA, Charache P, et al. Abundant Expression of EBER1 Small Nuclear RNA in Nasopharyngeal Carcinoma: A Morphologically Distinctive Target for Detection of Epstein-Barr Virus in Formalin-fixed Paraffin-embedded Carcinoma Specimens. Am J Pathol. 1991; 138(6): 1461-69.
18. Hanel P, Hummel M, Anagnostopoulos I, Stein H. Analysis of single EBER-positive and negative tumour cells in EBV-harbouring B-cell non-Hodgkin lymphomas. J Pathol. 2001; 195(3): 355-60.
19. Broussset P, Butet V, Chittal S, Selves J, Delsol G. Comparison of in-situ hybridization using different non isotopic probes for detection of EBV in nasopharyngeal carcinoma and IHC correlation with anti-latent membrane protein antibody. Lab Invest. 1992; 67(4): 457-64.
20. Murono S, Yoshizaki T, Tanaka S, Takeshita H, Park C, Furukawa M. Detection of Epstein-Barr virus in nasopharyngeal carcinoma by in situ hybridization and polymerase chain reaction. Laryngoscope. 1997; 107(4): 523-26.
21. Mäkitie AA, MacMillan C, Ho J, Shi W, Lee A, O’Sullivan B, et al. Loss of p16 expression has prognostic significance in human nasopharyngeal carcinoma. Clin Cancer Res. 2003; 9: 2177-84.