Immunohistochemical detection of ribonucleotide reductase in human breast tumors

T. Saeki, T. Takahashi, M. Okabe, A. Furuya, N. Hanai, K. Yamagami, K. Mandai, S. Moriwaki, H. Doihara, S. Takashima, D. S. Salomon

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

Ribonucleotide reductase (RNR) consists of two non-identical subunits, R1 and R2 and is one of the key enzymes involved in DNA biosynthesis. RNR activity is considerably higher in malignant tumors than in normal tissues in the rat suggesting that RNR may play an important role in the pathogenesis of human tumors. In order to obtain immunological reagents to study the localization and level of expression of RNR in various human tissues, a synthetic peptide containing sequences corresponding to the COOH-terminal region of the human R2 subunit was used to generate rat monoclonal antibodies. The generated rat monoclonal antibodies (IgG) inhibited RNR enzymatic activity purified from murine P388 leukemia cells. These antibodies were used to immunohistochemically examine the distribution of RNR in a small panel of 8 malignant and 4 benign human breast tumors. Positive immunostaining for RNR was observed in the cytoplasm of human breast carcinoma cells in which a specific 44 kDa specific band of R2 subunit was also detected by Western blot analysis. The immunostaining was blocked by preabsorption of the antibody with an excess amount of the synthetic peptide immunogen. In 8 of 8 breast carcinomas, positive immunostaining for the R2 subunit was observed whereas noninvolved, adjacent breast tissue showed no staining with this antibody. In addition, few of the benign breast lesions exhibited staining with this antibody. These data indicate that these antibodies can immunohistochemically detect RNR in frozen or formalin-fixed, paraffin- embedded tissues and that there is a differential expression of RNR between breast tumors and non-involved breast tissue. Immunohistochemical detection of RNR using these antibodies may therefore be useful for the diagnosis of human breast tumors.

Original languageEnglish
Pages (from-to)523-529
Number of pages7
JournalInternational journal of oncology
Volume6
Issue number3
Publication statusPublished - Jan 1 1995
Externally publishedYes

Keywords

  • human breast carcinomas
  • immunohistochemistry
  • ribonucleotide reductase (RNR)

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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