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Quantitation of estrogen receptor mRNA in breast carcinoma by branched DNA assay

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Abstract

A quantitative nucleic acid hybridization assay for determination of estrogen receptor (ER) mRNA in breast carcinoma is described. The assay, which is based on the branched DNA (bDNA) technology, requires 20 mg of tissue, is simple, highly specific, and reproducible, and correlates reasonably well with an established methodology (r=0.87). The assay has a dynamic range of 3 × 103 – 6 × 107 copies of ER mRNA per well. ER message as high as 2.5 × 106 copies per well could be detected in normal breast tissues. Thus a sensitivity of 3 × 103 ER copies per well was sufficient to analyze clinical specimens. In the present studies, accurate measurement of tissue weight enabled direct reporting of the ER mRNA values as the end point results. The bDNA assay provides a useful tool for the detection and quantitation of ER mRNA in research and routine clinical laboratories.

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References

  1. Holmes FA, Fritsche HA, Loewy JW, Geitner AM, Sutton RC, Buzdar AU, Hortobagyi GN: Measurement of estrogen and progesterone receptors in human breast tumors: Enzyme immunoassay versus binding assay. J Clin Oncol 8: 1025–1035, 1990

    Google Scholar 

  2. Gaffney EV, Sharmanov AT, Moody WE, Halpin DP, Blakemore WS, Elliot CB: Hormone receptor assays and their value in breast cancer therapy. Cancer Biotherapy 8: 17–28, 1993

    Google Scholar 

  3. Masood S: Estrogen and progesterone receptors in cytology: A comprehensive review. Diag Cytopathol 8: 475–491, 1992

    Google Scholar 

  4. Pasic R, Djulbegovic B, Wittliff JL: Comparison of sex steroid receptor determinations in human breast cancer by enzyme immunoassay and radioligand binding. J Clin Lab Anal 4: 430–436, 1990

    Google Scholar 

  5. Battifora H, Mehta P, Ahn C, Esteban JM: Estrogen receptor immunohistochemical assay in paraffin-embedded tissue. Appl Immunohistochem 1: 39–45, 1993

    Google Scholar 

  6. Daffada AA, Johnston SR, Nicholls J, Dowsett M: Detection of wild type and exon 5-deleted splice variant estrogen receptor (ER) mRNA in ER-positive and-negative breast cancer cell lines by reverse transcription polymerase chain reaction. J Mol Endocrinol 13: 265–273, 1994

    Google Scholar 

  7. Mies C: A simple, rapid method for isolating RNA from paraffin-embedded tissue for reverse transcription-polymerase chain reaction (RT-PCR), J Histochem Cytochem 42: 811–813, 1994

    Google Scholar 

  8. Koehorst SGA, Cox JJ, Donker GH, Lopes da Silva S, Burbach JPH, Thijssen JHH, Blankenstein MA: Functional analysis of an alternatively spliced estrogen receptor lacking exon 4 isolated from MCF-7 breast cancer cells and meningioma tissue. Mol Cell Endocrinol 101: 237–245, 1994

    Google Scholar 

  9. Leygue ER, Watson PH, Murphy LC: Estrogen receptor variants in normal human mammary tissue. J Natl Cancer Inst 88: 284–290, 1996

    Google Scholar 

  10. Graham DM, Jin L, Lloyd RV: Detection of estrogen receptor in paraffin-embedded sections of breast carcinoma by immunohistochemistry and in situ hybridization. Am J Surg Pathol 15: 475–485, 1991

    Google Scholar 

  11. Pachl C, Todd JA, Kern DG, Sheridan PJ, Fong SJ, Stempien M, Hoo B, Besemer D, Yeghiazarian T, Irvine B, Kolberg J, Kokka R, Neuwald P, Urdea MS: Rapid and precise quantification of HIV-1 RNA in plasma using a branched DNA signal amplification assay. J Acquir Immune Defic Syndr Hum Retrovirol 8: 446–454, 1995

    Google Scholar 

  12. Collins ML, Irvine B, Tyner D, Fine E, Zayati C, Chang CA, Horn T, Ahle D, Detmer J, Shen LP, Kolberg J, Bushnell S, Ho DD, Urdea MS: A branched DNA signal amplification assay for quantification of nucleic acid targets below 100 molecules/mL. Nucleic Acids Res 25: 2979–2984, 1997

    Google Scholar 

  13. Fuqua SAW, Chamness GC, McGuire WL: Estrogen receptor mutations in breast cancer. J Cell Biochem 51: 135–139, 1993

    Google Scholar 

  14. Greene GL, Gilna P, Waterfield M, Baker A, Hort Y, Shine J: Sequence and expression of human estrogen receptor complementary DNA. Science 231: 1150–1154, 1986

    Google Scholar 

  15. Collins ML, Zayati C, Detmer JJ, Daly B, Kolberg JA, Cha T, Irvine BD, Tucker J, Urdea MS: Preparation and characterization of RNA standards for use in quantitative branched DNA hybridization assays. Anal Biochem 226: 120–129, 1995

    Google Scholar 

  16. Terrault NA, Daily PJ, Ferrel L, Collins ML, Wilber JC, Urdea MS, Bhandari BN, Wright TL: Hepatitis C virus: Quantitation and distribution in liver. J Med Virol 51: 217–224, 1997

    Google Scholar 

  17. Dotzlaw H, Alkhalaf M, Murphy LC: Characterization of estrogen receptor variant mRNAs from human breast cancers. Mol Endocrinol 6: 773–785, 1992

    Google Scholar 

  18. Fuqua SAW, Wiltschke C, Castles C, Wolf D, Allred DC: A role for estrogen-receptor variants in endocrine resistance. Endocrine-Related Cancer 2: 19–25, 1995

    Google Scholar 

  19. Castles CG, Fuqua SAW, Koltz DM, Hill SM: Expression of a constitutively active estrogen receptor variant in the estrogen receptor negative BT-20 human breast cancer cell line. Cancer Res 53: 5934–5939, 1993

    Google Scholar 

  20. Murphy LC, Wang M, Coutt A, Dotzlaw H: Novel mutations in the estrogen receptor messenger RNA in human breast cancers. J Clin Endocrinol Metabol 81: 1420–1427, 1996

    Google Scholar 

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Authors and Affiliations

  1. Chiron Diagnostics, Alameda, California

    R.D. Nargessi, N.F. Khabbaz, X.M. Xu, M. Zamroud, J. Kolberg & M.L. Collins

  2. Emeryville, California, USA

    R.D. Nargessi, N.F. Khabbaz, X.M. Xu, M. Zamroud, J. Kolberg & M.L. Collins

Authors
  1. R.D. Nargessi
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  2. N.F. Khabbaz
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  3. X.M. Xu
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  4. M. Zamroud
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  5. J. Kolberg
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  6. M.L. Collins
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Nargessi, R., Khabbaz, N., Xu, X. et al. Quantitation of estrogen receptor mRNA in breast carcinoma by branched DNA assay. Breast Cancer Res Treat 50, 47–55 (1998). https://doi.org/10.1023/A:1006068811086

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  • DOI: https://doi.org/10.1023/A:1006068811086

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