Published in Cancer Detection and Prevention 1998; 22(Supplement 1).

Predicting disease progression of cervical intraepithelial neoplasia by monitoring high risk HPV oncoproteins with quantitative reverse transcriptase-PCR and novel immunocytochemical E6-E7 biomarkers

EB Holladay PhD, A Bhagavathiammai PhD, G Re PhD, D Nixon MD PhD, T Gue BS

AIM: To evaluate the relationship between quantities of viral transforming gene transcripts and cervical intraepithelial disease progression by monitoring oncogenic E6/E7 and HPV viral proteins with quantitative RT-PCR and novel E6 and E7 immunocytochemical biomarkers in paraffin-embedded biopsies. METHODS: To study the correlation between the progression pattern of the cervical intraepithelial lesions and the HPV mRNA transcription levels, the specimens were selected based on patients who had multiple retrospective biopsies ranging from cervical intraepithelial neoplasia (CIN) I to CIN III. RNA was isolated from formalin fixed paraffin embedded tissues and PCR amplification was performed to amplify specific cDNA sequences. A quantitative assay for determining the relative quantities of spliced E6 1 and E6 II transcripts for HPV-16 was performed. The results were correlated with immunocytochemical analysis of identical paraffin-embedded tissue using novel E6 and E7 monoclonal antibodies. RESULTS: Six of 6 patients with varying grades of CIN had detectable HPV 16 E6/E7 transcription. Specimens (n=12) from patients with confirmed histologic progression (CINI-CINII-CINIII) had increasing transcription levels. In one case, transcription patterns predicted dysplasia from a normal biopsy. Immunocytochemical analysis was less predictive than RT-PCR. In most cases, specimens positive for E7 oncoproteins had stronger signals than those positive for E6. CONCLUSIONS: Quantification of HPV-16 E6/E7 transcription may provide a prognostic tool for identifying women who are at increased risk for the progression of cervical intraepithelial neoplasia. Novel immunocytochemical monoclonal antibodies are currently less specific than RT-PCR for determining disease progression.

KEY WORDS: cervical intraepithelial neoplasia, E6/E7 transcription, Human Papillomavirus, Human Papillomavirus, immunocytochemistry, .

For more information, contact holladab@musc.edu

Paper presented at the International Symposium on the Impact of Biotechnology on Prediction, Prevention and Treatment of Cancer; Nice, France; October 24 - 27, 1998; in the section on Angiogenesis & Inhibition.

http://www.cancerprev.org/Journal/Issues/22/101/36/2398