HUMAN PAPILLOMAVIRUS IN VULVAR AND VAGINAL-CARCINOMA CELL-LINES

A number of reports associate human papillomavirus (HPV) with cervical cancer and cancer cell lines derived from this tumour type. Considera bly fewer reports have Focused on the role of HPV in carcinomas from o ther sites of female anogenital squamous epithelia. In this study we h ave tested for the presence of HPV in eight low-passage vulvar carcino ma cell lines and one extensively passaged cell line, A431. One cell l ine from a primary vaginal carcinoma was included. The presence of the HPV was evaluated by the polymerase chain reaction (PCR), by Southern blot analysis and by two-dimensional gel electrophoresis. General pri mer-mediated PCR was applied by using primers from the L1 region, E1 r egion and HPV 16 E7 region. Southern blot hybridisation was performed under low-stringency conditions (T-m = -35 degrees C) using a whole ge nomic HPV 6/16/18 probe mixture and under high stringency conditions ( T-m = -18 degrees C) with the whole genomic probes of HPV 16 and 33. H PV 16 E6-E7 mRNA was assessed by ribonuclease protection assay (RPA). HPV was found in only one vulvar carcinoma cell line, UM-SCV-6. The id entified type, HPV 16, was integrated in the cell genome and could be amplified with all primers used. Also E6-E7 transcripts were found in these cells. Five original tumour biopsies were available from the HPV -negative cell lines for in situ hybridisation. All these were HPV neg ative with both the HPV 6/16/18 screening probe mixture under low stri ngency and the HPV 16 probe under high stringency. The results indicat e that vulvar carcinoma cell lines contain HPV less frequently than ce rvical carcinoma cell lines and suggest that a significant proportion of vulvar carcinomas may evolve by an HPV-independent mechanism.