Tumor Formation in Nude Mice Inoculated with Cultured Human Epithelial Cells Co-Expressing Epstein-Barr Virus Latent Membrane Protein 1 and Bcl-2

Aims: To examine the tumor-forming in nude mice of human epithelial cells co-expressed Bcl-2 and EBV LMP-1 ability, the phenotype of tumor cells and their expression of oncogenes and tumor-suppressor genes. Methods: Following an in vivo tumorigenesis test in nude mice, the phenotype and growth properties of tumor cells were observed. Levels of expression of Bcl-2, and EBV LMP-1, and of onco- and tumor-suppressor proteins were detected by Western blot assay. Results: Human epithelial cells co-expressing Bcl-2 and EBV LMP-1 can form tumors in nude mice. Tumors appeared 48–65 days postinoculation in 6 of 10 nude mice tested but not in mice given Bcl-2-positive (0/5), LMP-1-positive (0/5) and RHEK-1 control (0/5) cells. Levels of c-myc protein were upregulated by LMP-1 but were not affected by Bcl-2 in this cell background; screening for other cellular oncogene and tumor suppressor gene products showed no change. Conclusion: The complementary effects of EBV LMP-1 and Bcl-2 in human epithelial cells resulted in tumor formation in nude mice but did not affect the expression of onco- and tumor suppressor proteins except for elevated c-myc. These findings suggest that LMP-1 and Bcl-2 can contribute together to the formation of EBV-associated epithelial cell tumors.