title: Detection of occult metastatic breast cancer cells in blood by a multimolecular marker assay: correlation with clinical stage of disease.
abstract: Currently, molecular markers offer the unique opportunity to identify occult metastasis in early stage cancer patients not otherwise detected with conventional staging techniques. To date, well-characterized molecular tumor markers to detect occult breast cancer cells in blood are limited. Because breast tumors are heterogeneous in tumor marker expression, we developed a "multimarker" reverse transcription-PCR assay combined with the highly sensitive electrochemiluminescence automated detection system. Breast cancer cell lines (n = 7), primary breast tumors (n = 25), and blood from normal donors (n = 40) and breast cancer patients [n = 65; American Joint Committee on Cancer (AJCC) stages I-IV] were assessed for four mRNA tumor markers: beta-human chorionic gonadotropin (beta-hCG), oncogene receptor (c-Met), beta 1-->4-N-acetylgalactosaminyl-transferase, and a tumor-associated antigen (MAGE-A3). None of the tumor markers were expressed in any normal donor bloods. Breast cancer cell lines and primary breast tumors expressed beta-hCG, c-Met, beta 1-->4-N-acetylgalactosaminyl-transferase, and MAGE-A3 mRNA. Of the 65 breast cancer patient blood samples assessed, 2, 3, 15, 49, and 31% expressed 4, 3, 2, 1, and 0 of the mRNA tumor markers, respectively. At least two markers were expressed in 20% of the blood specimens. The addition of a combination of markers enhanced detection of systemic metastasis by 32%. In patient blood samples, the MAGE-A3 marker correlated significantly with tumor size (P = 0.0004) and AJCC stage (P = 0.007). The combination of beta-hCG and MAGE-A3 mRNA markers correlated significantly with tumor size (P = 0.04), and the marker combination c-Met and MAGE-A3 showed a significant correlation with tumor size (P = 0.005) as well as AJCC stage (P = 0.018). A multimarker reverse transcription-PCR assay that correlates with known clinicopathological prognostic parameters may have potential clinical utility by monitoring tumor progression with a blood test.
authors.affiliation: Department of Molecular Oncology, John Wayne Cancer Institute, Santa Monica, California 90404, USA.
authors.forename: B
authors.initials: B
authors.lastname: Taback
authors.forename: A D
authors.initials: AD
authors.lastname: Chan
authors.forename: C T
authors.initials: CT
authors.lastname: Kuo
authors.forename: P J
authors.initials: PJ
authors.lastname: Bostick
authors.forename: H J
authors.initials: HJ
authors.lastname: Wang
authors.forename: A E
authors.initials: AE
authors.lastname: Giuliano
authors.forename: D S
authors.initials: DS
authors.lastname: Hoon
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000951:Antigens, Neoplasm / Q000096:biosynthesis / Q000097:blood / Q000235:genetics; D014408:Biomarkers, Tumor / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics; D001943:Breast Neoplasms / Q000097:blood* / Q000473:pathology*; D002822:Choriocarcinoma / Q000235:genetics / Q000378:metabolism; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000096:biosynthesis / Q000097:blood / Q000235:genetics; D005260:Female; D006801:Humans; D017350:N-Acetylgalactosaminyltransferases / Q000096:biosynthesis / Q000097:blood / Q000235:genetics; D009363:Neoplasm Proteins; D009367:Neoplasm Staging; D009360:Neoplastic Cells, Circulating / Q000378:metabolism / Q000473:pathology*; D019859:Proto-Oncogene Proteins c-met / Q000096:biosynthesis / Q000097:blood / Q000235:genetics; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D018997:Chorionic Gonadotropin, beta Subunit, Human; C072013:MAGEA3 protein, human; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D017350:N-Acetylgalactosaminyltransferases; C023220:(N-acetylneuraminyl)-galactosylglucosylceramide N-acetylgalactosaminyltransferase; D019859:Proto-Oncogene Proteins c-met