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      In vitro and in vivo Release of Soluble erbB‐2 Protein from Human Carcinoma Cells

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          Abstract

          An enzyme‐linked immunosorbent assay (ELISA) was developed to measure soluble erbB‐2 protein in culture supernatants of various human cell lines and sera of patients suffering from recurrent breast carcinoma. Soluble erbB‐2 protein was demonstrated in culture supernatants of cell lines that expressed high levels of erbB‐2 protein as shown by western blot analysis of cell lysates. Increased levels of the protein, 40‐ to 190‐fold higher than in healthy controls, were demonstrated in sera of 3 out of 12 patients with breast carcinomas. On immunohistological study of tumor tissues from 9 patients, high immune reaction with the anti‐ erbB‐2 protein antibody was observed in 2 cases. These were two of the three patients who had elevated levels of erbB‐2 protein in serum (a sample was not available from the third patient). These results raise the possibility that soluble erbB‐2 protein level in serum can be used as an indicator for spread of carcinomas that overexpress erbB‐2 protein.

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          Most cited references16

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          Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures.

          The use of avidin-biotin interaction in immunoenzymatic techniques provides a simple and sensitive method to localize antigens in formalin-fixed tissues. Among the several staining procedures available, the ABC method, which involves an application of biotin-labeled secondary antibody followed by the addition of avidin-biotin-peroxidase complex, gives a superior result when compared to the unlabeled antibody method. The availability of biotin-binding sites in the complex is created by the incubation of a relative excess of avidin with biotin-labeled peroxidase. During formation of the complex, avidin acts as a bridge between biotin-labeled peroxidase molecules; and biotin-labeled peroxidase molecules, which contains several biotin moieties, serve as a link between the avidin molecules. Consequently, a "lattice" complex containing several peroxidase molecules is likely formed. Binding of this complex to the biotin moieties associated with secondary antibody results in a high staining intensity.
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            Tyrosine kinase receptor with extensive homology to EGF receptor shares chromosomal location with neu oncogene.

            A novel potential cell surface receptor of the tyrosine kinase gene family has been identified and characterized by molecular cloning. Its primary sequence is very similar to that of the human epidermal growth factor receptor and the v-erbB oncogene product; the chromosomal location of the gene for this protein is coincident with the neu oncogene, which suggests that the two genes may be identical.
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              The product of the human c-erbB-2 gene: a 185-kilodalton glycoprotein with tyrosine kinase activity.

              Antibodies were raised against a synthetic peptide corresponding to 14 amino acid residues at the COOH-terminus of a protein deduced from the human c-erbB-2 nucleotide sequence. These antibodies immunoprecipitated a 185-kilodalton glycoprotein from MKN-7 adenocarcinoma cells. Incubation of the immunoprecipitates with (gamma-32P)ATP resulted in the phosphorylation of this protein on tyrosine residues. These results indicate that the human c-erbB-2 gene product is the 185-kilodalton glycoprotein that is associated with tyrosine kinase activity. Although the c-erbB-2 protein was predicted to encode a protein very similar to epidermal growth factor (EGF) receptor, EGF did not stimulate this kinase activity either in vivo or in vitro.
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                Author and article information

                Journal
                Jpn J Cancer Res
                Jpn. J. Cancer Res
                10.1111/(ISSN)1349-7006a
                CAS
                Japanese Journal of Cancer Research : Gann
                Blackwell Publishing Ltd (Oxford, UK )
                0910-5050
                1876-4673
                May 1990
                : 81
                : 5 ( doiID: 10.1111/cas.1990.81.issue-5 )
                : 489-494
                Affiliations
                [ 1 ]Departments of Pathology, Institute of Medical Science, University of Tokyo, 4–6‐1 Shirokanedai, Minato‐ku, Tokyo 108
                [ 2 ]Research Institute, Nichirei Co., 1–52‐14 Kumegawa, Higashi‐Murayama, Tokyo 189
                [ 3 ]Department of Clinical Oncology, Cancer Institute Hospital and Division of Clinical Chemotherapy, Cancer Chemotherapy Center
                [ 4 ]Departments of Oncology, Institute of Medical Science, University of Tokyo, 4–6‐1 Shirokanedai, Minato‐ku, Tokyo 108
                [ 5 ]Department of Pathology, Cancer Institute, Japanese Foundation for Cancer Research, 1–37‐1 Kami‐Ikebukuro, Toshima‐ku, Tokyo 170
                Author notes
                [*] [* ]To whom correspondence should be sent.
                Article
                CAE489
                10.1111/j.1349-7006.1990.tb02596.x
                5918074
                1974247
                abd68794-c32f-4b76-951a-6c58baeef2a8
                History
                Page count
                References: 19, Pages: 6
                Categories
                Article
                Custom metadata
                2.0
                May 1990
                Converter:WILEY_ML3GV2_TO_NLMPMC version:4.6.9 mode:remove_FC converted:04.11.2015

                receptor,oncogene,carcinoma,membrane protein,breast
                receptor, oncogene, carcinoma, membrane protein, breast

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