Characterization of EBV-Infected Epithelial Cell Lines from Gastric Cancer-Bearing Tissues
The long-term goal of our study is to explore how Epstein-Barr virus (EBV) associates with the pathogenesis of gastric carcinoma. The first goal is to establish EBV-positive epithelial cell lines from EBV-infected gastric carcinoma tissues and to characte
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T. SAIRENJl I ,6, M, TAHMA2, M, KANAMORIl, N, TAKASAKAl,
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . .
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2 "Spontaneous" Establishment of EBV-Positive Gastric Epithelial Cell Lines 2.1 GT38 and GT39 Cell Lines . . . . . . . . . . 2.2 Chromosomal Analysis
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3 3.1 3.2 3.3 3.4 3.5
EBV Infection in GT38 and GT39 Cell Lines. Latency Type . . . . Genotype of EBV .. . Spontaneous EBV Reactivation. EBV Reactivation with TPA .. Production of Infectious Virus .
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4 4.1 4.2 4.3 4.4
Nitric Oxide Downregulates EBV Reactivation. iNOS Gene Expression in GT38 and GT39 Cell Lines. EBV Reactivation by a Competitive Inhibitor of NOS, L-NMMA Effect of TPA on EBV Reactivation and iNOS mRNA Expression. Effect of L-NMMA and SNAP, a NO Donor on TPA-Induced EBV Reactivation.
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Loss of EBV DNA Copy in GT38 and GT39 Cell Lines
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6 Oncogenic Potential . . . . . . 6.1 Cell Growth in Soft Agar. . . 6.2 Tumorigenesis in SCID Mice . 7
Conclusions.
References .
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1 Introduction The long-term goal of our study is to explore how Epstein-Barr virus (EBV) associates with the pathogenesis of gastric carcinoma. The first goal is to establish
I Department of Biosignaling, School of Life Science, Faculty of Medicine, Tottori University, Yon ago 683-8503, Japan 2 Central Clinical Laboratory, Teikyo University School of Medicine, Tokyo 173-0003, Japan 3 Department of Second Surgery, Teikyo University School of Medicine, Tokyo 173-0003, Japan 4 Department of Clinical Pathology, Teikyo University School of Medicine, Tokyo 173-0003, Japan 5 First Department of Pathology, Faculty of Medicine, Tottori University, Yonago 683-8503, Japan 6 Department of Pathology, National Institute ofInfectious Diseases, Tokyo 162-0052, Japan
K. Takada (ed.), Epstein-Barr Virus and Human Cancer © Springer-Verlag Berlin Heidelberg 2001
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T. Sairenji et al.
EBV -positive epithelial cell lines from EBV-infected gastric carcinoma tissues and to characterize the cell lines and EBV infection in the cells. EBV, a ubiquitous human herpesvirus with oncogenic potential, is predominantly associated with the infection of two target tissues in vivo: (1) B lymphocytes, where the infection is largely nonproductive, and (2) the epithelium, in which virus replication occurs (RICKINSON and KIEFF 1996). Both target tissues are susceptible to EBV-associated malignant change, leading to tumors of B-cell origin, such as Burkitt's lymphoma, or of epithelial-cell origin, such as nasopharyngeal carcinoma (NPC) (PATHMANATHAN et al. 1995). Recently, EBV has also emerged as an etiologic agent implicated in gastric carcinoma (MIN et al. 1991; SHIBATA et al. 1991; SHIBATA and WEISS 1992; TOKUNAGA et al. 1993a,b; FUKAYAMA et al. 1994; IMAI et al. 1994; OHFUJI et al. 1996; IWASAKI et al. 1998). EBV has been found in most cases of rare gastric lymphoepithelioma-like carcinomas (MIN et al. 1991; SHIBATA et al. 1991; OHFUJI et al. 1996) and a small but significant proportion of common gastric aden
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