cDNA and protein characterization of human MAGE‐10

• Published in: International journal of cancer 1999-09, Vol.82 (6), p.901-907
• Main Authors: Rimoldi, Donata, Salvi, Suzanne, Reed, Darryl, Coulie, Pierre, Jongeneel, Victor C., De Plaen, Etienne, Brasseur, Francis, Rodriguez, Anne‐Marie, Boon, Thierry, Cerottini, Jean‐Charles
• Format: Article
• Language: English
• Subjects: Antibodies, Monoclonal; Antigens, Neoplasm - genetics; Biological and medical sciences; Blotting, Northern; Cloning, Molecular; Dermatology; DNA, Complementary; Female; Gene Library; Host-tumor relations. Immunology. Biological markers; Humans; Introns; Male; Medical sciences; Melanoma - genetics; Neoplasm Proteins - chemistry; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Organ Specificity; Polymerase Chain Reaction; Protein Biosynthesis; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; Transcription, Genetic; Transfection; Tumors; Tumors of the skin and soft tissue. Premalignant lesions
• ISSN: 0020-7136; 1097-0215
• DOI: 10.1002/(SICI)1097-0215(19990909)82:6<901::AID-IJC21>3.0.CO;2-X

MAGE genes are frequently expressed in several types of human malignancy and code for antigens recognized by cytotoxic T lymphocytes. We have previously described a monoclonal antibody (MAb), named 6C1, that recognizes the MAGE‐1 protein and cross‐reacts with a 72‐kDa protein present in lysates of melanoma cells such as MZ2‐MEL. To identify this protein, we have screened an expression library prepared from MZ2‐MEL cells. Several clones that encoded a protein recognized by antibody 6C1 contained a sequence identical to that of MAGE‐10, another member of the MAGE‐A gene family. Full‐length MAGE‐10 cDNA clones, obtained after screening additional cDNA melanoma libraries, were found to be approximately 2.5 kb in length. In vitro translation and transient transfection experiments indicated that MAGE‐10 codes for a protein of approximately 72 kDa. This product was recognized by MAb 6C1 as well as by a polyclonal serum raised against a MAGE‐10 peptide, thus demonstrating its identity with MAGE‐10. Analysis of MAGE‐10 mRNA by RT‐PCR confirmed its presence in testis and placenta but not in other normal tissues. Expression of MAGE‐10 in melanoma tumors was found to parallel that of MAGE‐1. Western blot analysis with the polyclonal anti‐MAGE‐10 antibody showed the presence of MAGE‐10 in lysates of purified trophoblast cells. Immuno‐cytochemistry of cultured melanoma cells indicated that MAGE‐10 is a nuclear protein. Int. J. Cancer 82:901–907, 1999. © 1999 Wiley‐Liss, Inc.