Identification of integrated Epstein-Barr virus in nasopharyngeal carcinoma using pulse field gel electrophoresis.
Integrated Epstein-Barr virus (EBV) was found in 4 of 17 pathologically diagnosed, EBV-positive nasopharyngeal carcinoma (NPC) biopsy samples using pulse field gel electrophoresis (PFGE). The analyses were carried out after digestion of biopsy DNA with rare restriction enzymes such as PacI, which does not cleave EBV due to the absence of the recognition sequence, and ClaI. Employing this technique, EBV integration in IB4, through EcoI, and Namalwa through the terminal repeats, was confirmed. We also established that integration of EBV in AW Ramos was through the terminal repeats. The Raji cell line was also found to harbour integrated virus in addition to episomal DNA. However, the site of integration could not be confirmed, since as the Raji DNA appeared to be heavily methylated and could not be cleaved with CPG rare cutters. As in the Raji cell line, the DNA of EBV in nasopharyngeal-carcinoma (NPC) biopsies also appeared to be methylated.
Kripalani-Joshi
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Institute of Molecular and Cell Biology, National University of Singapore.
Law
H Y
HY
D000818:Animals; D001402:B-Lymphocytes / Q000382:microbiology; D001706:Biopsy; D002144:Callithrix; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004279:DNA, Viral / Q000032:analysis; D016521:Electrophoresis, Gel, Pulsed-Field; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000382:microbiology* / Q000473:pathology; D014407:Tumor Cells, Cultured
D004279:DNA, Viral; D004262:DNA Restriction Enzymes