TY - JOUR
T1 - A transcribed gene, containing a variable number of tandem repeats, codes for a human epithelial tumor antigen
T2 - cDNA cloning, expression of the transfected gene and over‐expression in breast cancer tissue
AU - HAREUVENI, Mara
AU - TSARFATY, Ilan
AU - ZARETSKY, Joseph
AU - KOTKES, Phillip
AU - HOREV, Judith
AU - ZRIHAN, Sheila
AU - WEISS, Mordechai
AU - GREEN, Stephen
AU - LATHE, Richard
AU - KEYDAR, Iafa
AU - WRESCHNER, Daniel H.
PY - 1990/5
Y1 - 1990/5
N2 - A monoclonal antibody, H23, that specifically recognizes a breast‐tumor‐associated antigen, was used to isolate a cDNA insert that codes for the antigenic epitope. Nucleotide sequencing of this cDNA, as well as a longer 850‐bp cDNA insert, shows that they are composed of 60‐bp (G + C)‐rich tandem repeating units. The coding strand was determined and codes for a proline‐rich 20‐amino‐acid repeat motif. A comparison of the highly conserved repeat unit with the deduced flanking amino acid sequences demonstrates conservation of specific subregions of the repeat consensus within the flanking amino acids. Hybridization of the 60‐bp cDNA probe with RNAs extracted from a variety of primary and metastatic human tumors yields relatively high levels of hybrid with the breast carcinomas, as compared to lower hybrid levels with RNAs from other epithelial tumors. RNA extracted from breast tissue adjacent to the tumor or from benign breast tumors, demonstrates low or undetectable levels of hybridization. Probing Southern blots with the 60‐bp repeat shows that the tumor antigen is highly polymorphic and contains a variable number of tandem repeats (VNTRs). The VNTR nature of the gene was confirmed by probing Southern blots with unique genomic sequences that are physically linked to an isolated gene fragment that also contains the tandem repeat array. Mouse cells transfected with this gene fragement produce tumor antigen that is readily detected by H23 monoclonal antibodies. The allelic forms seen in 10 different primary human tumors demonstrate 100% concordance with the various mRNA species expressed. These studies are extended to the protein forms detected by immunoblot analyses that show both a correlation of the expressed tumor antigen species with the allelic forms as well as significantly increased expression in breast cancer tissue. The above studies unequivocally establish the over‐expression of a VNTR gene coding for an epithelial tumor antigen in human breast cancer tissue.
AB - A monoclonal antibody, H23, that specifically recognizes a breast‐tumor‐associated antigen, was used to isolate a cDNA insert that codes for the antigenic epitope. Nucleotide sequencing of this cDNA, as well as a longer 850‐bp cDNA insert, shows that they are composed of 60‐bp (G + C)‐rich tandem repeating units. The coding strand was determined and codes for a proline‐rich 20‐amino‐acid repeat motif. A comparison of the highly conserved repeat unit with the deduced flanking amino acid sequences demonstrates conservation of specific subregions of the repeat consensus within the flanking amino acids. Hybridization of the 60‐bp cDNA probe with RNAs extracted from a variety of primary and metastatic human tumors yields relatively high levels of hybrid with the breast carcinomas, as compared to lower hybrid levels with RNAs from other epithelial tumors. RNA extracted from breast tissue adjacent to the tumor or from benign breast tumors, demonstrates low or undetectable levels of hybridization. Probing Southern blots with the 60‐bp repeat shows that the tumor antigen is highly polymorphic and contains a variable number of tandem repeats (VNTRs). The VNTR nature of the gene was confirmed by probing Southern blots with unique genomic sequences that are physically linked to an isolated gene fragment that also contains the tandem repeat array. Mouse cells transfected with this gene fragement produce tumor antigen that is readily detected by H23 monoclonal antibodies. The allelic forms seen in 10 different primary human tumors demonstrate 100% concordance with the various mRNA species expressed. These studies are extended to the protein forms detected by immunoblot analyses that show both a correlation of the expressed tumor antigen species with the allelic forms as well as significantly increased expression in breast cancer tissue. The above studies unequivocally establish the over‐expression of a VNTR gene coding for an epithelial tumor antigen in human breast cancer tissue.
UR - http://www.scopus.com/inward/record.url?scp=0025337106&partnerID=8YFLogxK
U2 - 10.1111/j.1432-1033.1990.tb15512.x
DO - 10.1111/j.1432-1033.1990.tb15512.x
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C2 - 2112460
AN - SCOPUS:0025337106
SN - 0014-2956
VL - 189
SP - 475
EP - 486
JO - European Journal of Biochemistry
JF - European Journal of Biochemistry
IS - 3
ER -