Molecular strategy for detecting metastatic cancers with use of multiple tumor-specific MAGE-A genes.
BACKGROUND
The human melanoma-associated antigen family A (MAGE-A) has high specificity and expression in various malignancies, but individual family members are expressed at low frequency in any one particular type of cancer. We therefore developed a method to detect mRNAs from multiple MAGE-A genes in a single reaction.


METHODS
Universal MAGE-A (uMAGE-A) primers and probe were designed to reverse-transcribe, amplify, and detect by electrochemiluminescence (ECL) MAGE-A mRNAs on the Origen Analyzer. The assay was performed on total RNA of melanoma (n = 9 cell lines and 24 tumors), breast cancer (n = 7 and 26), and colorectal cancer (CRC; n = 5 and 12). We also evaluated blood from melanoma (n = 50), breast cancer (n = 16), and CRC (n = 21) patients.


RESULTS
The uMAGE-A mRNA was detectable in 0.01-1 ng of cell line RNA. The identity of the uMAGE-A cDNA products was confirmed by sequencing and polyacrylamide gel electrophoresis. The uMAGE-A assay increased detection of melanoma, breast cancer, and CRC tumor by 13%, 31%, and 25%, respectively, compared with a MAGE-A1 assay, and by 17%, 19%, and 25%, respectively, compared with a MAGE-A3 assay. The uMAGE-A assay detected circulating tumor cells in the blood of melanoma (24%), breast cancer (25%), and CRC (29%) patients.


CONCLUSIONS
The uMAGE-A reverse transcription-PCR/ECL assay provides a practical and sensitive approach for detection of various metastatic cancers in tissues and blood.
Department of Molecular Oncology, John Wayne Cancer Clinic, Division Gastrointestinal Surgery, and Joyce Eisenberg Keefer Breast Center, John Wayne Cancer Institute, Santa Monica, CA 90404, USA.
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Miyashiro
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Kuo
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Huynh
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Iida
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Morton
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Bilchik
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Giuliano
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Hoon
D000951:Antigens, Neoplasm / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000097:blood / Q000378:metabolism*; D001943:Breast Neoplasms / Q000097:blood / Q000235:genetics* / Q000473:pathology; D015179:Colorectal Neoplasms / Q000097:blood / Q000235:genetics* / Q000473:pathology; D005260:Female; D006801:Humans; D008163:Luminescent Measurements; D008545:Melanoma / Q000097:blood / Q000235:genetics* / Q000473:pathology; D058950:Melanoma-Specific Antigens; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000097:blood / Q000235:genetics* / Q000378:metabolism; D009360:Neoplastic Cells, Circulating; D012333:RNA, Messenger / Q000097:blood / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; C409589:MAGEA12 protein, human; C072013:MAGEA3 protein, human; D058950:Melanoma-Specific Antigens; D009363:Neoplasm Proteins; D012333:RNA, Messenger