HUMAN PAPILLOMAVIRUS TYPE-18 E6-ASTERISK MESSENGER-RNA IN PRIMARY TUMORS AND PELVIC LYMPH-NODES OF HUNGARIAN PATIENTS WITH SQUAMOUS CERVICAL-CANCER

Seven biopsy specimens from squamous-cell carcinomas of the uterine ce rvix were examined by RT-PCR for human-papilloma-virus(HPV)-specific t ranscripts. With our HPV18-transcription-specific primer pair (5' nts 127-149; 3' nts 587-607), all 7 were shown to contain one strong viral mRNA signal from the early 6/early 7 open reading frames (E6/E7 ORFs) . Sequence analysis of the cloned PCR product proved that the transcri pt was generated by splicing out an intron in E6 from nucleotides 233 to 416, thereby corresponding to the HPV18 E6 spliced mRNA. Nine out of 9 metastatic and 5 of 7 histologically negative lymph nodes from th e same patients were also found to be positive for the same mRNA trans cript. However, 4 HPV18 unrelated primary tumors and the connected reg ional pelvic lymph nodes (3 metastatic, 7 histologically negative) wer e negative for the HPV18 E6 mRNA. Cytokeratin signals indicating tumo r cells of epithelial origin were detected in 7 out of the 9 transcrip t-positive lymph nodes with histological signs of metastasis and in 2 out of the 5 transcript-positive histologically negative lymph nodes. This suggests that the dispersion of the epithelial monoclonal tumor c ells was lymphogenic in origin. (C) 1992 Wiley-Liss, Inc.