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.
Miyashiro
I
I
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.
Kuo
C
C
Huynh
K
K
Iida
A
A
Morton
D
D
Bilchik
A
A
Giuliano
A
A
Hoon
D S
DS
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