| Abstract: |
The prostate cancer HERV-K gag-related NGO-Pr-54 antigen was identified by SEREX analysis using autologous patient serum. NGO-Pr-54 mRNA was observed to be faintly expressed in normal prostate and strongly expressed in a variety of cancers, including ovarian cancer (5/8), prostate cancer (6/9), and leukemia (5/14). A phage plaque assay showed that a strong reaction was constantly observed with clone ZH042 in which the 5' end of NGO-Pr-54 is deleted, suggesting that it contained the sequence coding for the protein product. A TI-35 mAb was produced using a recombinant protein (438 aa) deduced from the sequence of ZH042. Transfection of clone ZH042 into 293T cells resulted in the production of an approximately 50-kDa molecule visualized by Western blotting. Natural production of the molecule was confirmed in a SK-MEL-23 melanoma cell line. An indirect immunofluorescence assay showed that NGO-Pr-54 protein was expressed on the cell surface as well as in the cytoplasm. Cell surface expression was confirmed by flow cytometry using the TI-35 mAb. The antibody response against NGO-Pr-54 was observed in patients with bladder (5.1%), liver (4.1%), lung (3.4%), ovarian (5.6%), and prostate (4.2%) cancer, as well as with malignant melanoma (13.2%). |
| Keywords: |
protein expression; aged; aged, 80 and over; leukemia; unclassified drug; gene sequence; genetics; cancer patient; flow cytometry; molecular genetics; ovarian neoplasms; mouse; animal; metabolism; animals; mice; melanoma; ovary cancer; gene expression; neoplasm proteins; cell line; cancer screening; transfection; cercopithecus aethiops; cos cells; tumor antigen; enzyme linked immunosorbent assay; mice, inbred balb c; monoclonal antibody; prostate cancer; prostatic neoplasms; gene expression regulation; molecular cloning; cloning, molecular; fluorescent antibody technique; gene expression regulation, neoplastic; immunology; antibodies, monoclonal; prostate; amino acid sequence; molecular sequence data; bagg albino mouse; genetic transfection; antigens, neoplasm; messenger rna; immunogenicity; ovary tumor; prostate tumor; recombinant proteins; recombinant protein; tumor protein; western blotting; cell strain cos1; cercopithecus; melanoma cell; autoantigen; immunofluorescence test; tumor gene; molecular weight; serum; antiserum; fluorescent antibody technique, indirect; cell surface; gag protein; gag; herv-k; serex; ngo pr 54 protein, human; ngo-pr-54 protein, human; human endogenous retrovirus; serological recombinant cdna expression; endogenous retrovirus; endogenous retroviruses; gene products, gag; immune sera
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