PMID- 10521804
OWN - NLM
STAT- MEDLINE
DCOM- 19991119
LR  - 20190708
IS  - 0020-7136 (Print)
IS  - 0020-7136 (Linking)
VI  - 83
IP  - 5
DP  - 1999 Nov 26
TI  - Quantitative evaluation of the expression of MAGE genes in tumors by limiting
      dilution of cDNA libraries.
PG  - 664-9
AB  - The MAGE-A genes are expressed in tumor cells but not in healthy tissues, except 
      in male germ line cells and in placenta. They encode tumor-specific antigens
      recognized by autologous cytolytic T lymphocytes (CTLs). On the basis of
      semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR)
      assays, 6 of the 12 members of the MAGE-A family, including MAGE-A1, were
      previously reported to have a high level of expression in tumors, whereas 5 other
      members, including MAGE-A10, were expressed at a much lower level, deemed to be
      insufficient for CTL recognition. However, analysis with antibodies has shown
      that some melanoma cell lines contain equivalent amounts of MAGE-A1 and MAGE-A10 
      proteins. This discrepancy appeared to be due to the low efficacy of the primers 
      that had been used for the previous MAGE-A10 RT-PCR assays. This led us to
      develop a method that is independent of the efficacy of the PCR primers to
      evaluate MAGE-A gene expression. cDNA libraries from tumor cell lines were
      introduced into bacteria, of which 200 pools of about 500 bacteria were
      maintained in microcultures. The frequencies of the MAGE-A cDNA clones in each
      library were evaluated by performing PCR assays on each of these pools. The
      abundance of MAGE-A10 cDNAs was found to be similar to that of MAGE-A1 in 3 of
      the libraries that were analyzed, including 2 with high expression (1/6,400),
      confirming that MAGE-A10 is expressed at a high level. MAGE-A2, A3, A4, A6 and
      A12 cDNAs were also confirmed often to be present at a frequency of more than
      1/10,000, a level of expression that should suffice for recognition of antigenic 
      peptides encoded by these genes by cytolytic T cells. The remaining MAGE genes
      are either not expressed in tumors or are expressed at a very low level, with the
      exception of MAGE-A8 and 11, which show high expression in a very small number of
      tumors. This method also allowed us to isolate 5 MAGE-A cDNAs that we had not
      obtained previously, enabling us to delineate the exons in the sequences of genes
      MAGE-A5, A8, A9, A10 and A11.
CI  - Copyright 1999 Wiley-Liss, Inc.
FAU - Serrano, A
AU  - Serrano A
AD  - Ludwig Institute for Cancer Research, Brussels Branch, Brussels, Belgium.
FAU - Lethe, B
AU  - Lethe B
FAU - Delroisse, J M
AU  - Delroisse JM
FAU - Lurquin, C
AU  - Lurquin C
FAU - De Plaen, E
AU  - De Plaen E
FAU - Brasseur, F
AU  - Brasseur F
FAU - Rimoldi, D
AU  - Rimoldi D
FAU - Boon, T
AU  - Boon T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Int J Cancer
JT  - International journal of cancer
JID - 0042124
RN  - 0 (Antigens, Neoplasm)
RN  - 0 (MAGE-A10 antigen)
RN  - 0 (MAGEA1 protein, human)
RN  - 0 (Melanoma-Specific Antigens)
RN  - 0 (Neoplasm Proteins)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Antigens, Neoplasm/*genetics/isolation & purification/metabolism
MH  - Chromosome Mapping
MH  - *Gene Expression Regulation, Neoplastic
MH  - *Gene Library
MH  - Humans
MH  - Male
MH  - Melanoma-Specific Antigens
MH  - Neoplasm Proteins/*genetics/isolation & purification/metabolism
MH  - Placenta/metabolism
MH  - RNA, Messenger/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Testis/metabolism
MH  - Tumor Cells, Cultured
EDAT- 1999/10/16 00:00
MHDA- 1999/10/16 00:01
CRDT- 1999/10/16 00:00
PHST- 1999/10/16 00:00 [pubmed]
PHST- 1999/10/16 00:01 [medline]
PHST- 1999/10/16 00:00 [entrez]
AID - 10.1002/(SICI)1097-0215(19991126)83:5<664::AID-IJC16>3.0.CO;2-V [pii]
AID - 10.1002/(sici)1097-0215(19991126)83:5<664::aid-ijc16>3.0.co;2-v [doi]
PST - ppublish
SO  - Int J Cancer. 1999 Nov 26;83(5):664-9. doi:
      10.1002/(sici)1097-0215(19991126)83:5<664::aid-ijc16>3.0.co;2-v.