Determination of EBV genotypes and EBNA1 oncogene variants among breast cancer patients in Peshawar, Pakistan

Authors

  • Ayesha Khattak Institute of Zoological Sciences, University of Peshawar Pakistan
  • Sanaullah Khan Institute of Zoological Sciences, University of Peshawar Pakistan
  • Fatima Nauman Institute of Zoological Sciences, University of Peshawar Pakistan
  • Itrat Sabeen Institute of Zoological Sciences, University of Peshawar Pakistan
  • Muhammad Bar Khan Institute of Zoological Sciences, University of Peshawar Pakistan

DOI:

https://doi.org/10.59736/IJP.23.04.987

Keywords:

Breast Cancer, Epstein Barr Virus, Genotypes, Nuclear Antigen

Abstract

Background: Epstein-Barr virus (EBV) is a common DNA virus linked to several human cancers, including breast cancer (BC). Specific EBV oncogenes, like EBNA1, are also implicated in the process of oncogenesis. This study aimed to investigate the prevalence of different EBV genotypes and EBNA1 oncogene variants in BC patients in Peshawar, Pakistan.

Methodology: This study was conducted in the Institute of zoological sciences, University of Peshawar, Khyber Pakhtunkhwa.  Tumor biopsies and demographic data were collected from 75 BC patients. DNA was extracted from the samples and EBV genotypes were detected using polymerase chain reaction (PCR) with specific primers. The EBNA1 oncogene was also amplified via PCR and sequenced using Sanger's sequencing method. The resulting sequences were analyzed to determine EBNA1 variants. All data were analyzed using descriptive statistics, primarily percentages and chi square test where applicable.

Results: EBV was detected in 40% of the study participants. All positive samples were identified as EBV genotype-1. The most prevalent EBNA1 type found in these patients was the prototype P-Threonine. The study population was predominantly post-menopausal (63%), with most tumors being malignant (93%) and of grade II (69%). Among the malignant cases, 63% were hormone receptor (HR) positive, 11% were HER2-positive, and 26% were triple-negative breast cancer (TNBC).

Conclusion: EBV type-1 and EBNA1 P-Threonine are present in breast cancer patients in Peshawar, Pakistan. This study provides valuable insight into the molecular characteristics of EBV in breast cancer within this specific population.

References

Cao, Y., Xie, L., Shi, F. et al. Targeting the signaling in Epstein–Barr virus-associated diseases: mechanism, regulation, and clinical study. Sig Transduct Target Ther. 2021 Jan 12;6(1):15.

Shechter O, Sausen DG, Gallo ES, Dahari H, Borenstein R. Epstein–Barr virus epithelial-associated malignancies: exploring pathologies and current treatments. Int J Mol Sci. 2022;23(22):14389.

Gómez-Archila JD, Arellano-Galindo J, Palacios-Reyes C, Espinosa-García AM, Alonso-Themann PG, Xicohtencatl-Cortes J, et al. Epstein-Barr virus as a promoter of tumorigenesis in the tumor microenvironment of breast cancer. Int J Mol Med. 2023; 52 (2): 72.

Huang X, Zhang M & Zhang Z. The role of LMP1 in Epstein-Barr virus-associated gastric cancer. Curr Cancer Drug Targets. 2024;24(2):127–41.

Epstein MA, Achong BG, Barr YM. Virus Particles in Cultured Lymphoblasts from Burkitt’s Lymphoma. Lancet. 1964; 1: 702–703.

Mashaly M, Ghorab D, Hegazy M, Abdelkhalek M, Gaballa K, Elzehery R. Association between Epstein-Barr Virus Gene Polymorphism and Risk among Egyptian Females. Asian Pac J Cancer Prev. 2022; 23 (2): 641-50.

Sharifpour C, Makvandi M, Samarbafzadeh A, Talaei-Zadeh A, Ranjbari N, Nisi N, et al. Frequency of Epstein–Barr Virus DNA in Formalin-Fixed Paraffin-Embedded 16 Tissue of Patients with Ductal Breast Carcinoma. Asian Pac J Cancer Prev. 2019; 20 (3): 687-92.

Kang MS, Kieff E. Epstein–Barr virus latent genes. Exp Mol Med. 2015;47: e131

Zhang X, Li Z, Peng Q, Liu C, Wu Y, Wen Y, et al. Epstein–Barr virus suppresses N6-methyladenosine modification of TLR9 to promote immune evasion. J Biol Chem. 2024;300(5):107226.

Smith DW, Bill Sugde B. Potential Cellular Functions of Epstein-Barr Nuclear Antigen 1 (EBNA1) of Epstein-Barr Virus. Viruses. 2013; 5: 226-40.

Frappier L. Ebna1. 2025.

Jiang L, Xie C, Lung HL, Lo KW, Law GL, Mak NK & Wong KL. EBNA1-targeted inhibitors: Novel approaches for the treatment of Epstein-Barr virus-associated cancers. Theranostics. 2018; 8 (19): 5307.

Sun L, Zhao Z, Liu S, Liu X, Sun Z & Luo B. Sequence variation analysis of Epstein-Barr virus nuclear antigen 1 gene in the virus associated lymphomas of Northern China. PloS one. 2015; 10 (10): e0140529.

Globalcancerobservatory.2022.https,/gco.iarc.who.int/media/globocan/factsheets/populations/586-pakistan-fact-sheet.pdf (accessed on 21/04/2024)

National Breast Cancer Coalition (NBCC), Facts and figures 2022. https, //www.stopbreastcancer.org/information-center/facts-figures/ (accessed on 21.08/2024)

Zaheer S, Shah N, Maqbool SA, Soomro NM. Estimates of past and future time trends in age-specific breast cancer incidence among women in Karachi, Pakistan: 2004–2025. BMC Public Health. 2019; 19 (1): 1001.

Bibi R, Maneewat K, Sangchan H, Sae-sia W. Persistent post-surgical pain and its relationship to health-related quality of life in Pakistani women after breast cancer surgery. Belitung Nurs J. 2022; 8 (3): 258-65.

BangashNA.DawnE-PaperPakistan. 2023. https://www.dawn.com/news/1782011 (accessed on 21/08/ 2024).

Khan NU, Khan BM, Azam I & Hayat A. Current Situation of Breast Cancer in Pakistan and the Dire Need of Early Diagnosis. Current Trends in OMICS. 2024; 4 (2): 1-20.

Tabatabaie FH, Hosseini SY, Hashemi SMA, Safaie A, Sarvari J. A preliminary sequence analysis of the Epstein-Barr virus nuclear antigen 1 (EBNA1) carboxy-terminal region in cervical and ovarian cancers. Iran J Pathol. 2023; 18 (1): 24.

Zhang Y, Ji Y, Liu S, Li J, Wu J, Jin Q, et al. Global burden of female breast cancer: new estimates in 2022, temporal trends and future projections up to 2050. J Natl Cancer Cent. 2025;5(3):287.

Mundo L, Leoncini L, Accardi-Gheit R. Epstein–Barr virus infection in cancer. Cancers (Basel). 2023;15(18):4659.

Niedobitek G, Meru N, Delecluse HJ. Epstein–Barr virus infection and human malignancies. Int J Exp Pathol. 2001; 82:149–70

Michaels E, Worthington RO, Rusiecki J. Breast cancer: risk assessment, screening, and primary prevention. Med Clin North Am. 2023; 107 (2): 271–84

Gatti V, Bongiorno-Borbone L, Fierro C, Annicchiarico-Petruzzelli M, Melino G, Peschiaroli A. p63 at the crossroads between stemness and metastasis in breast cancer. Int J Mol Sci. 2019;20(11):2683.

Abdalah UE, Shalabi MG, Elderdery AY, Abdelbagi A, Mohamed H, Babker AMA, et al. Association of Epstein–Barr virus detection with histopathologic-prognostic findings among Sudanese breast cancer patients. Int J Pharm Res. 2020;12.

Ballard AJ. Epstein-Barr virus is associated with aggressive subtypes of invasive ductal carcinoma of breast (Her2+/ER- and triple negative) and with nuclear expression of NF-κB p50. Arch Breast Cancer. 2018 Jun 8;68–75.

Ma CX, Suman VJ, Sanati S, et al. Endocrine-Sensitive Disease Rate in Postmenopausal Patients with Estrogen Receptor–Rich/ERBB2-Negative Breast Cancer Receiving Neoadjuvant Anastrozole, Fulvestrant, or Their Combination: A Phase 3 Randomized Clinical Trial. JAMA Oncol. 2024;10(3):362–71. doi:10.1001/jamaoncol.2023.6038

Breast Cancer UK. Menopause and breast cancer. 2024. Available from: https://www.breastcanceruk.org.uk. Accessed 9 Mar 2025.

Smatti MK, Al-Sadeq DW, Ali NH, Pintus G, Abou-Saleh H and Nasrallah GK. Epstein–Barr Virus Epidemiology, Serology, and Genetic Variability of LMP-1 Oncogene Among Healthy Population: An Update. Front. Oncol. 2018; 211.

Mofrad MG, Kazeminezhad B, Faghihloo E. Prevalence of Epstein–Barr virus (EBV) in Iranian Breast Carcinoma Patients. Asian Pac J Cancer Prev. 2020;21.

Mazouni C, Fina F, Romain S, Ouafik L, Bonnier P, Brandone JM. Martin PM. Epstein–Barr virus as a marker of biological aggressiveness in breast cancer. Br. J. Cancer. 2011; 104: 332–7.

Shahi V, Agarwal P, Qayoom S, Kumar V, Tewari S, Raghuvanshi S, et al. Detection of EBNA-1, early antigen, and LMP1 in breast cancer of Northern India. Asian Pac J Cancer Prev. 2022;23(11):3717.

Banko AV, Lazarevic IB, Karalic DZ, Djukic VB, Cupic MD, Stevanovic G, Jovanovic TP. Sequence analysis of Epstein–Barr virus EBNA1 gene in nasopharyngeal carcinoma. Med Microbiol Immunol. 2019; 208:81–8.

Chang KL, Chen YY, Chen WG, Hayashi K, Bacchi C, Bacchi M, Weiss LM. EBNA-1 Gene Sequences in Brazilian and American Patients with Hodgkin's Disease. Blood. 1999; 94 (1): 244-50.

Wang LU, Tian WD, Xu X, Nie B, Lu J, Liu X, et al. EBNA1 protein induces epithelial–mesenchymal transition in nasopharyngeal carcinoma cells. Cancer. 2014;120(3):363–72.

Yu J, Wang Y, Wang H, Wei Z, Pei Y. Decoding Critical Targets and Signaling Pathways in EBV-Mediated Diseases Using Large Language Models. Viruses. 2024; 16: 1660.

Ayee R, Ofori MEO, Wright E, Quaye O. Epstein–Barr virus-associated lymphomas and epithelial cancers in humans. J Cancer. 2020;11(7):1737.

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Published

2025-12-31

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Original article

How to Cite

1.
Khattak A, Khan S, Nauman F, Sabeen I, Khan MB. Determination of EBV genotypes and EBNA1 oncogene variants among breast cancer patients in Peshawar, Pakistan. Int J Pathol [Internet]. 2025 Dec. 31 [cited 2025 Dec. 31];23(4):236-43. Available from: https://jpathology.com/index.php/OJS/article/view/987

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