title: Quantitative real-time RT-PCR detection of breast cancer micrometastasis using a multigene marker panel.
abstract: 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 x 10(5)-fold above normal mean expression, whereas PIP was overexpressed from 30- to 2.2 x 10(6)-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 mam, PIP, CK19, mamB, muc1 and CEA can be applied as a panel for detection of metastatic and occult micrometastatic disease.
authors: Department of Surgery, Medical University of South Carolina, Charleston, SC 29425, USA. mitasm@musc.edu
authors: M
authors: M
authors: Mitas
authors: K
authors: K
authors: Mikhitarian
authors: C
authors: C
authors: Walters
authors: P L
authors: PL
authors: Baron
authors: B M
authors: BM
authors: Elliott
authors: T E
authors: TE
authors: Brothers
authors: J G
authors: JG
authors: Robison
authors: J S
authors: JS
authors: Metcalf
authors: Y Y
authors: YY
authors: Palesch
authors: Z
authors: Z
authors: Zhang
authors: W E
authors: WE
authors: Gillanders
authors: D J
authors: DJ
authors: Cole
mesh_terms: D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000235:genetics; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D003199:Computer Systems; D017931:DNA Primers; D018076:DNA, Complementary / Q000032:analysis; D005260:Female; D006801:Humans; D008198:Lymph Nodes / Q000378:metabolism; D008207:Lymphatic Metastasis / Q000235:genetics; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis / Q000175:diagnosis; D011379:Prognosis; D012333:RNA, Messenger / Q000096:biosynthesis; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D014408:Biomarkers, Tumor; D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger