Production of EBV genome-chip for screening viral infection and Ggne expressions in human tumors
aDepartment of Microbiology and Immunology, Chung Shan Medical University, Taichung, Taiwan, bDepartment of Microbiology and Immunology, Chang-Gung University, Taoyuan, Taiwan
Aim: Microarray is a powerful tool for genomic studies. Based on its high capacity and specificity, this pilot study was designed to test the feasibility of using EBV (Epstein-Barr virus)-chip to screen human tumor specimens for the infection of EBV, the virus has been associated to B-cell lymphomas/leukemia, nasopharyngeal carcinomas, and the growing numbers of cancer cases from other organs. Methods: Seventy-one EBV DNA fragments of 1-3 kbp in length were amplified by PCR using specific primers designed for the entire EBV genome. These amplified products were used to fabricate EBV-chip, followed by testing the specificity with mRNA prepared from P3HR1 cell line harboring EBV and tumors tissues from various organs. Results: The strong gene expression signals were detected by EBV-chips hybridized to the biotin-labeled cDNA that was converted from mRNA of P3HR1 cells activated to allow EBV replication, but not from the cells containing the latent viral genome. Forty-four different tumor tissues were also assayed with EBV-chips and revealed interesting viral gene expression patterns. Conclusions: The production of EBV genome-chip is very useful to screen various tumors for the infection of EBV that may lead to the insights into the tumorigenic mechanism associated to the virus in the following investigations.
Paper presented at the International Symposium on Predictive Oncology and Intervention Strategies; Nice, France; February 7 - 10, 2004; in poster session 791 (Viral infections).