Epstein-Barr virus infection to Epstein-Barr virus-negative nasopharyngeal carcinoma cell line TW03 enhances its tumorigenicity

Norihiro Teramoto, Akihiko Maeda, Keita Kobayashi, Kazuhiko Hayashi, Takashi Oka, Kiyoshi Takahashi, Kenzo Takada, Georg Klein, Tadaatsu Akagi

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Abstract

Almost all nasopharyngeal carcinomas (NPCs) are infected by Epstein-Barr virus (EBV), but most ex vivo NPC cells lose EBV genomes during passages. In this study, an EBV-negative NPC cell line, TW03, established from EBV- carrying NPC was reinfected with EBV by cocultivation with irradiated Akata cells carrying recombinant EBV containing a neomycin-resistant gene. The reinfected EBV (+) TW03 cells expressed EBERs and EBNA1, but not EBNA2, lytic proteins (ZEBRA and EA-D), or LMP1. They had an epithelial appearance similar to that of EBV (-) TW03 cells. The doubling times of EBV (+) and EBV () TW03 cells were almost identical. However, the EBV (+) TW03 cells formed larger colonies with ragged contours in anchorage-independent cultures. An in vitro invasion assay showed that EBV (+) TW03 cells had a higher invasive activity than EBV () TW03 cells (p <0.01). Both EBV () and EBV (+) TW03 cells formed poorly differentiated squamous cell carcinomas in SCID and nude mice. EBV (+) TW03 cells showed a higher tumorigenicity to nude mice (12 of 13) than EBV (- ) TW03 cells (1 of 9) (p <0.001). In the severe combined immunodeficiency (SCID) tumors of EBV (+) TW03 cells, not all of the tumor cells were EBER-1 positive. EBER-1 was more frequently detected in the peripheral regions and daughter nodules of the tumors than in the central areas. The microdissection polymerase chain reaction showed that the EBER-1 -negative TW03 cells in the EBV (+) TW03 SCID tumors lost EBV genomes. EBER-1-negative cells showed as high a rate of Ki-67 positivity as EBER-1-positive cells, indicating that the former were proliferating rather than dead or dying. In horny pearls, keratinizing cells were ZEBRA-positive and EBER-negative. Loss of EBV genomes was not associated with squamous differentiation. These data indicated that reinfection of EBV promotes the tumorigenicity of EBV (-) TW03 cells by enhancing the invading activity.

Original languageEnglish
Pages (from-to)303-312
Number of pages10
JournalLaboratory Investigation
Volume80
Issue number3
Publication statusPublished - Mar 2000

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Epstein-Barr Virus Infections
Human Herpesvirus 4
Cell Line
Severe Combined Immunodeficiency
Nasopharyngeal carcinoma
Genome
Nude Mice
Neoplasms

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Teramoto, N., Maeda, A., Kobayashi, K., Hayashi, K., Oka, T., Takahashi, K., ... Akagi, T. (2000). Epstein-Barr virus infection to Epstein-Barr virus-negative nasopharyngeal carcinoma cell line TW03 enhances its tumorigenicity. Laboratory Investigation, 80(3), 303-312.

Epstein-Barr virus infection to Epstein-Barr virus-negative nasopharyngeal carcinoma cell line TW03 enhances its tumorigenicity. / Teramoto, Norihiro; Maeda, Akihiko; Kobayashi, Keita; Hayashi, Kazuhiko; Oka, Takashi; Takahashi, Kiyoshi; Takada, Kenzo; Klein, Georg; Akagi, Tadaatsu.

In: Laboratory Investigation, Vol. 80, No. 3, 03.2000, p. 303-312.

Research output: Contribution to journalArticle

Teramoto, N, Maeda, A, Kobayashi, K, Hayashi, K, Oka, T, Takahashi, K, Takada, K, Klein, G & Akagi, T 2000, 'Epstein-Barr virus infection to Epstein-Barr virus-negative nasopharyngeal carcinoma cell line TW03 enhances its tumorigenicity', Laboratory Investigation, vol. 80, no. 3, pp. 303-312.
Teramoto, Norihiro ; Maeda, Akihiko ; Kobayashi, Keita ; Hayashi, Kazuhiko ; Oka, Takashi ; Takahashi, Kiyoshi ; Takada, Kenzo ; Klein, Georg ; Akagi, Tadaatsu. / Epstein-Barr virus infection to Epstein-Barr virus-negative nasopharyngeal carcinoma cell line TW03 enhances its tumorigenicity. In: Laboratory Investigation. 2000 ; Vol. 80, No. 3. pp. 303-312.
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abstract = "Almost all nasopharyngeal carcinomas (NPCs) are infected by Epstein-Barr virus (EBV), but most ex vivo NPC cells lose EBV genomes during passages. In this study, an EBV-negative NPC cell line, TW03, established from EBV- carrying NPC was reinfected with EBV by cocultivation with irradiated Akata cells carrying recombinant EBV containing a neomycin-resistant gene. The reinfected EBV (+) TW03 cells expressed EBERs and EBNA1, but not EBNA2, lytic proteins (ZEBRA and EA-D), or LMP1. They had an epithelial appearance similar to that of EBV (-) TW03 cells. The doubling times of EBV (+) and EBV () TW03 cells were almost identical. However, the EBV (+) TW03 cells formed larger colonies with ragged contours in anchorage-independent cultures. An in vitro invasion assay showed that EBV (+) TW03 cells had a higher invasive activity than EBV () TW03 cells (p <0.01). Both EBV () and EBV (+) TW03 cells formed poorly differentiated squamous cell carcinomas in SCID and nude mice. EBV (+) TW03 cells showed a higher tumorigenicity to nude mice (12 of 13) than EBV (- ) TW03 cells (1 of 9) (p <0.001). In the severe combined immunodeficiency (SCID) tumors of EBV (+) TW03 cells, not all of the tumor cells were EBER-1 positive. EBER-1 was more frequently detected in the peripheral regions and daughter nodules of the tumors than in the central areas. The microdissection polymerase chain reaction showed that the EBER-1 -negative TW03 cells in the EBV (+) TW03 SCID tumors lost EBV genomes. EBER-1-negative cells showed as high a rate of Ki-67 positivity as EBER-1-positive cells, indicating that the former were proliferating rather than dead or dying. In horny pearls, keratinizing cells were ZEBRA-positive and EBER-negative. Loss of EBV genomes was not associated with squamous differentiation. These data indicated that reinfection of EBV promotes the tumorigenicity of EBV (-) TW03 cells by enhancing the invading activity.",
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