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Suppression of Epstein-Barr nuclear antigen 1 (EBNA1) by RNA interference inhibits proliferation of EBV-positive Burkitt’s lymphoma cells

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Abstract

Purpose: Epstein-Barr virus (EBV) is associated with the development of several lymphoid and epithelial malignancies, including Burkitt’s lymphoma. The EBV latent protein, EBV Nuclear Antigen 1 (EBNA1), is detectable in almost all types of EBV-associated tumors and is essential for replication and maintenance of the latent episome of EBV. We here examined whether the RNA interference (RNAi) technique could be employed to suppress expression of EBNA1 in EBV-positive Burkitt’s lymphoma cells. Methods: A Raji cell line expressing small hairpin RNAs (shRNAs) against EBNA1 was established and EBNA1 mRNA level was determined by real-time RT-PCR analysis. We investigated the effects of EBNA1 silence on lymphoma cell growth and cell cycle progression. Results: Transfection of an EBNA1 RNAi plasmid resulted in substantial loss of EBNA1 mRNA and significantly inhibited proliferation of Raji cells relative to the control plasmid case. Suppression of EBNA1 was also associated with downregulation of EBV oncogene EBNA2, a decreased PCNA labeling index and increased G0/G1 fraction in cell cycle analysis. Conclusions: These findings point to potential therapeutic applications for vector-mediated siRNA delivery to control EBV-associated malignant disorders.

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Acknowledgements

We would like to thank Associate Professor Hiroyuki Kishi for advice on flow cytometry analysis of transfection efficiency and Tokimasa Kumada and Hideki Hatta for their expert technical support. We are also grateful to Associate Professor Hideto Yonekura and Dr Hui Li (Kanazawa University, Japan) for helpful discussions and Drs Fumihiro Tomoda and Hua-Chuan Zheng for their assistance with the statistical analysis. This work was supported in part by a Grant-in-aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Authors and Affiliations

  1. Department of Pathology, School of Medicine, Toyama Medical and Pharmaceutical University, 2630 Sugitani, 930-0194, Toyama, Japan

    Mei Hong, Yoshihiro Murai, Tomohiko Kutsuna, Hiroyuki Takahashi, Kazuhiro Nomoto, Chun-Mei Cheng, Shin Ishizawa, Koichi Tsuneyama & Yasuo Takano

  2. Department of Radiological Sciences, School of Medicine, Toyama Medical and Pharmaceutical University, 2630 Sugitani, 930-0194, Toyama, Japan

    Qing-Li Zhao & Ryohei Ogawa

  3. School of Life Sciences, Queensland University of Technology, 4000, Brisbane, Australia

    Brian V. Harmon

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  1. Mei Hong
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  2. Yoshihiro Murai
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Correspondence to Yasuo Takano.

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Hong, M., Murai, Y., Kutsuna, T. et al. Suppression of Epstein-Barr nuclear antigen 1 (EBNA1) by RNA interference inhibits proliferation of EBV-positive Burkitt’s lymphoma cells. J Cancer Res Clin Oncol 132, 1–8 (2006). https://doi.org/10.1007/s00432-005-0036-x

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  • DOI: https://doi.org/10.1007/s00432-005-0036-x

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