Quantitative real-time RT-PCR detection of breast cancer micrometastasis using a multigene marker panel

Michael Mitas, Kaidi Mikhitarian, Christian Walters, Paul L. Baron, Bruce M. Elliott, Thomas E. Brothers, Jay G. Robison, John S. Metcalf, Yuko Y. Palesch, Zhen Zhang, William E. Gillanders, David J. Cole

Research output: Contribution to journalArticlepeer-review

188 Scopus citations


Real-time RT-PCR is a relatively new technology that uses an online fluorescence detection system to determine gene expression levels. It has the potential to significantly improve detection of breast cancer metastasis by virtue of its exquisite sensitivity, high throughput capacity and quantitative readout system. To assess the utility of this technology in breast cancer staging, we determined the relative expression levels of 12 cancer-associated genes (mam, PIP, mamB, CEA, CK19, VEGF, erbB2, muc1, c-myc, p97, vim and Ki67) in 51 negative-control normal lymph nodes and in 17 histopathology-positive ALNs. We then performed a receiver operating characteristic (ROC) curve analysis to determine the sensitivity and specificity levels of each gene. Areas under the ROC curve indicated that the most accurate diagnostic markers were mam (99.6%), PIP (93.3%), CK19 (91.0%), mamB (87.9%), muc1 (81.5%) and CEA (79.4.0%). mam was overexpressed in 16 of 17 lymph nodes known to contain metastatic breast cancer at levels ranging from 22- to 2.8 × 105-fold above normal mean expression, whereas PIP was overexpressed from 30- to 2.2 × 106-fold above normal in 13 lymph nodes. Real-time RT-PCR analysis of pathology-negative LN from breast cancer patients revealed evidence of overexpression of PIP (6 nodes), mam (3 nodes) and CEA (1 node) in 8 of 21 nodes (38%). Our results provide evidence that mare, PIP, CK19, mamB, muc1 and CEA can be applied as a panel for detection of metastatic and occult micrometastatic disease.

Original languageEnglish
Pages (from-to)162-171
Number of pages10
JournalInternational Journal of Cancer
Issue number2
StatePublished - Jul 15 2001


  • Axillary lymph node
  • Breast cancer genes
  • Diagnostic
  • ROC analysis
  • SYBR Green I chemistry
  • Sentinel lymph node


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