DETECTION OF HUMAN-PAPILLOMAVIRUS-16 TRANSCRIPTIONAL ACTIVITY IN CERVICAL INTRAEPITHELIAL NEOPLASIA GRADE-III LESIONS AND CERVICAL CARCINOMAS BY NESTED REVERSE TRANSCRIPTION-POLYMERASE CHAIN-REACTION AND IN SITE HYBRIDIZATION

Continued expression of the oncogenes E6 and E7 of human papilloma vir us ''high risk'' type 16 (HPV16) initiates neoplastic transformation a nd maintenance of the malignant phenotype in cervical carcinoma cells. The transcriptional activity of the HPV16 E6/E7 oncogenes was investi gated in HPV16-containing cervical cell lines, cervical carcinomas, ac id cervical intraepithelial neoplasia grade III lesions using the tech niques of reverse transcription-polymerase chain reaction (RT-PCR), So uthern blotting, and in situ hybridization. To facilitate detection of the full-length HPV16 E6/E7 oncogene transcript and its characteristi c splice products E6I and E6*II in cervical tissues, a nested RT-PCR (nRT-PCR) assay was designed. Specific detection of HPV E6/E7 oncogene transcripts in clinical specimens was found to be improved by nRT-PCR , being as sensitive as the combination of conventional RT-PCR and sub sequent Southern blot hybridization. Regarding the progression of prem alignant lesions to cervical cancer, detection of the HPV transcriptio nal activity by nRT-PCR may provide additional information for risk ev aluations. Moreover, improvements in the amplification of HPV oncogene transcripts may also be advantageous for monitoring the activity of H PV before and after transcript-targeted gene therapy of cervical cance r.