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Detection of the metastatic potential of blood-borne and immunomagnetically enriched epithelial cells by quantitative erbB-2 RT-PCR

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A quantitative competitive RT-PCR targeting the specific mRNA of c-erbB-2 is described. It is well known that in many cancers the c-erbB-2 gene dosage is elevated and/or the mRNA is overexpressed leading to a poor prognosis. The recently established method to enrich epithelial cells from peripheral blood made it reasonable and logical to develop a sophisticated competitive RT-PCR method to estimate the c-erbB-2 mRNA load of such cells. By this enrichment method it was discovered that cancer cells can be either isolated as single cells or clusters from patients' blood. The plausibility for their metastatic potential is derived from the measurement of the c-erbB-2 mRNA expression. A SKBR3 cell model suggested that the c-erbB-2 mRNA load is between 168 and 336 molecules per cell. Results of the quantitative competitive RT-PCR method can be used as a rational to determine the efficiency of anti-oncogenic therapies and as a criterion in adjuvant therapy decision.

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References

  1. Clark WH, 1991, Tumor progression and the nature of cancer. Br J Cancer, 64, 631–44.

    Google Scholar 

  2. Moll R, Franke WW, Schiller DL, Geiger B and Krepler R, 1982, The catalog of human cytokeratins: patterns of expression in normal epithelia, tumours and cultured cells. Cell, 31, 11–32.

    Google Scholar 

  3. Griwatz C, Brandt B, Assmann G and Zänker KS, 1995, An immunological enrichment method for epithelial cells from peripheral blood. J Immunol Meth, 183, 251–65.

    Google Scholar 

  4. Hirokoshi T, Danenberg KD, Stadtbauer et al. 1992, Quantification of thymidylate synthase, dihydrofolate reductase, and DT-diaphorase gene expression in human tumors using the polymerase chain reaction. Cancer Res, 52, 108–16.

    Google Scholar 

  5. Benz CC, Brandt BH and Zänker KS, 1995, Gene diagnostics provide new insights into breast cancer prognosis and therapy. Gene, 159, 3–7.

    Google Scholar 

  6. Brandt B, Vogt U, Harms F, Jacksich C, Assmann G and Zänker KS, 1993, Breast cancer prognosis: identification of high-risk patients by erbB-2 and c-myc oncogene amplification. Proc Amer Soc Clin Oncol, 12, 237.

    Google Scholar 

  7. Brandt B, Vogt U, et al. 1994, Estimation of average gene copy number of erbB-1, erbB-2, and erbB-3 for prognosis in breast cancer. Proc Amer Assoc Cancer Res, 35, 212.

    Google Scholar 

  8. Semba K, Kamata N, Toyoshima K and Yamamoto T, 1985, A v-erbB-related protooncogene, c-erbB-2, is distinct from the c-erbB-1/epidermal growth factor-receptor gene and is amplified in a human salivary gland adenocarcinoma. Proc Natl Acad Sci USA, 82, 6497–501.

    Google Scholar 

  9. Ishii S, Xu YH, Stratton RH, Roe BA, Merlino GT and Pastan I, 1985, Characterisation and sequence of the promoter region of the human epidermal growth factor receptor gene. Proc Natl Acad Sci USA, 82, 4920–4.

    Google Scholar 

  10. Brandt B, Vogt U, Harms F, Bosse U, Zänker KS and Assmann G, 1995, Double-differential PCR for gene dosage estimation of erbB oncogenes in benign and cancer tissues, and comparison to cellular DNA content. Gene, 159, 29–34.

    Google Scholar 

  11. Fidler IJ, 1990, Critical factors in the biology of human cancer metastasis: twenty-eight. G.H.A. Clowes Memorial Award Lecture. Cancer Res, 50, 6130.

    Google Scholar 

  12. Friedl P, Noble PB, Walton PA, et al. 1995, Migration of coordinated cell clusters in mesenchymal and epithelial cancer explants in vitro. Cancer Res, 55, 4557–60.

    Google Scholar 

  13. Slamon DJ, Godolphin W, Jones LA, et al. 1989, Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science, 244, 707–12.

    Google Scholar 

  14. Sarkar FH, Visscher DW and Crissman JD, 1993, Quantitative analysis of Her-2/neu (ERBB2) gene expression using reverse transcriptase polymerase chain reaction. Diagn Mol Pathol, 2, 210–18.

    Google Scholar 

  15. Wels W, Moritz D, Schmidt M, Jeschke M, Hynes NE and Groner B, 1995, Biotechnological and gene therapeutic strategies in cancer treatment. Gene, 159, 73–80.

    Google Scholar 

  16. Morrison C and Gannon F, 1994, The impact of the plateau phase on quantitative PCR. Biochim Biophys Acta, 1219, 493–8.

    Google Scholar 

  17. Datta YH, Adams PT, Drobyski WR, Ethier SP, Terry VH and Roth MS, 1994, Sensitive detection of occult breast cancer by the reverse-transcriptase polymerase chain reaction. J Clin Onco, 12, 475–82.

    Google Scholar 

  18. Neumeier M, Gerhard M and Wagener C, 1995, Diagnosis of micrometastases by the amplification of tissue-specific genes. Gene, 159, 43–7.

    Google Scholar 

  19. Burchill S, Bradbiry M, Pittman K, Southgate L, Smith B and Selby P, 1995, Detection of epithelial cancer cells in peripheral blood by reverse transcriptase-polymearse chain reaction. Br J Cancer, 71, 278–83.

    Google Scholar 

  20. Moll R, Schiller DL and Franke WW, 1990, Identification of protein IT of the intestinal cytoskeleton as anovel type I cytokeratin with unusual properties and expression patterns. J Cell Biol, 111, 576–80.

    Google Scholar 

  21. Gerhard M, Juhl H, Kalthoff H, Schreiber HW, Wagener C and Neumeier M, 1994, Specific detection of carcinoembryonic antigen-expressing tumor cells in bone marrow aspirates by polymerase chain reaction. J Clin Oncol, 14, 235–8.

    Google Scholar 

  22. Tsai, CM, Chang KT, Perng RP, et al. 1993, Correlation of intrinsic chemoresistance of non-small cell lung cancer cell lines with Her-2/neu gene expression but not with ras gene mutations. J Natl Cancer Inst, 85, 897–901.

    Google Scholar 

  23. Schlimok G, Funke I, Pantel K, et al. 1991, Micrometastatic tumour cells in bone marrow of patients with gastric cancer: methodological aspects of detection and prognostic significance. Eur J Cancer, 27, 1461–5.

    Google Scholar 

  24. Pantel K, Schlimok G, Braun S, et al. 1993, Differential expression of proliferation-associated molecules in individual micrometastatic carcinoma cells. J Natl Cancer Inst, 85, 1419–23.

    Google Scholar 

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Brandt, B., Griwatz, C., Heidl, S. et al. Detection of the metastatic potential of blood-borne and immunomagnetically enriched epithelial cells by quantitative erbB-2 RT-PCR. Clin Exp Metast 14, 399–408 (1996). https://doi.org/10.1007/BF00123399

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  • DOI: https://doi.org/10.1007/BF00123399

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