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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 Situ Hybridization |
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Hans-Christoph
Selinka, Karl Sotlar, Karin Klingel, Martina Sauter, Reinhard Kandolf, and
Burkhard Bultmann |
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Institute
for Pathology, Department of Molecular Pathology, University of Tubingen,
Tubingen, Germany |
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Continued expression of
the oncogenes E6 and E7 of human papilloma virus "high risk'' type
16 (HPV16) initiates neoplastic transformation and maintenance of the malignant
phenotype in cervical carcinoma cells. The transcriptional activity of the
HPV16 E6/E7 oncogenes was investigated in HPV16-containing cervical cell
lines, cervical carcinomas, and cervical intraepithelial neoplasia grade
III lesions using the techniques of reverse transcription-polymerase chain
reaction (RT-PCR), Southern blotting, and in situ hybridization. To facilitate
detection of the full-length HPV16 E6/E7 oncogene transcript and its characteristic
splice products E6*I 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 subsequent Southern blot hybridization.
Regarding the progression of premalignant lesions to cervical cancer, detection
of the HPV transcriptional activity by nRT-PCR may provide additional information
for risk evaluations. Moreover, improvements in the amplification of HPV
oncogene transcripts may also be advantageous for monitoring the activity
of HPV before and after transcript-targeted gene therapy of cervical cancer. |
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