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Isolation of the Epstein-Barr virus in scirrhous gastric cancer by efficiency-monitored representational difference
1First Department of Internal Medicine, School of Medicine, Sapporo Medical University, Sapporo, Japan.
Abstract:
We applied an improved representational difference analysis (RDA), efficiency-monitored RDA (EM-RDA), to analyze the genetic alterations of scirrhous gastric cancer. Clinical samples of this cancer are difficult to analyze genetically because of the large number of normal cells in the stroma. The entire 3rd difference product obtained from EM-RDA was subcloned. This is equivalent to the scirrhous gastric cancer DNA with the normal gastric tissue DNA subtracted from it. The difference clones, which were confirmed by Southern blot as being derived from the cancer, were sequenced. As a result, two difference clones were identified as DNA fragments of the Epstein-Barr virus (EBV). The existence of EBV in the scirrhous gastric cancer cells was proved by EBV-encoded small RNA 1 in situ hybridization. EM-RDA made it possible to detect the first case of EBV-related gastric cancer in a young woman.