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Oncogene amplification in archival ovarian carcinoma detected by fluorescent differential polymerase chain reaction - a routine analytical approach
  1. M.W. Beckmann*,
  2. H.X. An*,,
  3. K. Köhrer,
  4. M. Finken-Eigen,
  5. W. Schröder,§,
  6. H.G. Schnürch* and
  7. H.G. Bender*
  1. *Department of Obstetrics and Gynecology and †Centre for Biological and Medical Research (BMFZ), Heinrich-Heine University, Dusseldorf, Germany; ‡Tongii Hospital, Tongii Medical University, Wuhan, China; §Department of Obstetrics and Gynecology, University of Aachen, Aachen, Germany
  1. Address for correspondence: Dr M. W. Beckmann, Universitäts-Frauenklinik, Moorenstrasse 5, 40225 Düsseldorf, Germany.


For quantitative determination of gene amplification of oncogenes involved in ovarian cancer development we have established a rapid, nonradioactive approach. Determination of c-erbB-2 and c-myc gene amplification in archival ovarian carcinoma specimens was based on differential polymerase chain reaction (dPCR) and fluorescent DNA technique.c-erb B-2 or c-myc sequences were amplified by PCR, in which one of each primer pair was fluorescently labeled, simultaneously with a reference gene (γ-IFN) as internal single copy gene control. PCR products were separated by polyacrylamide gel electrophoresis with an automated DNA sequencer (A.L.F.™, Pharmacia, Freiburg, Germany) and directly quantified after laser activation and emission scanning using appropriate software (Fragment Manager™, Pharmacia, Freiburg, Germany). This fluorescent differential PCR (fdPCR) method was used for quantitative determination of c-erbB-2 and c-myc gene amplification in 79 formalin-fixed, paraffin-embedded epithelial ovarian carcinoma tissues and 15 benign ovarian tissues. c-erbB-2 gene amplification was found in 18 (22%) and c-myc gene amplification in 13 (17%) of these carcinomas, but could not be detected in benign ovarian tissues. Single oncogene amplification correlated significantly with higher stage and higher grade (P<0.05). Patients with either c-erbB-2 orc-myc gene amplification or both had significantly shorter relapse-free survival and overall survival. The fdPCR assay is a valuable tool for determination of oncogene amplification even in archival tumor tissue. More detailed information about individual tumor biology of the primary tumor and metastasis as well as criteria for additional therapeutic decisions may be acquired by this analytical approach.

  • fdPCR
  • gene amplification
  • ovarian cancer
  • predictor
  • prognosticator

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