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Laboratoire de Génétique Moléculaire, Faculté des Sciences Pharmaceutiques et Biologiques de Paris, 4 Avenue de l'Observatoire, F-75006 Paris, France.
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Laboratoire d'Oncogénétique, Centre
René Huguenin, F-92211 St-Cloud, France.
a Author for correspondence. Fax 33 1 44 07 17 54; e-mail mvidaud{at}teaser.fr
Background: Gene amplification/overexpression of ERBB2 (HER2, neu) is a major event in human breast tumorigenesis. ERBB2-based therapeutic agents and ERBB2-specific gene therapy are under development. These new perspectives call for a sensitive and accurate method to screen breast cancer patients for ERBB2 alterations.
Methods: We have developed and validated a real-time quantitative reverse transcription (RT)-PCR assay, based on fluorescent TaqMan methodology, to quantify ERBB2 gene expression at the mRNA level in breast tumors. This recently developed method of nucleic acid quantification in homogeneous solutions has the potential for a wide dynamic range, interlaboratory agreement, and high-throughput capacity without tedious post-PCR processing. The ERBB2 mRNA signal was normalized to the signal for TATA box-binding protein mRNA.
Results: The dynamic range was >1000-fold. The relationship
between Ct and log starting concentration was linear
(r2 
0.99). The mean (SD) normalized expression
of ERBB2 in healthy breast tissue was 0.95 (0.37).
Overexpression (>5 SD above mean for healthy breast) of the
ERBB2 gene was observed (at 3.2- to 135-fold) in
23 (17%) of 134 breast tumor RNA samples. As expected,
ERBB2 overexpression was present in all tumors with
ERBB2 gene amplification but was uncommon and at a low
ratio (<5) in breast cancers without gene amplification.
Conclusions: This new simple, rapid, semi-automated assay is a major alternative to fluorescence in situ hybridization and immunochemistry for gene alteration analysis in human tumors and may be a powerful tool for large randomized, prospective cooperative group trials and to support future ERBB2-based biological and gene therapy approaches.© 1999 American Association for Clinical Chemistry
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G. M. Yousef, A. Scorilas, L. G. Kyriakopoulou, L. Rendl, M. Diamandis, R. Ponzone, N. Biglia, M. Giai, R. Roagna, P. Sismondi, et al. Human Kallikrein Gene 5 (KLK5) Expression by Quantitative PCR: An Independent Indicator of Poor Prognosis in Breast Cancer Clin. Chem., August 1, 2002; 48(8): 1241 - 1250. [Abstract] [Full Text] [PDF]
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D. G. Thomas, T. J. Giordano, D. Sanders, J. S. Biermann, and L. Baker Absence of HER2/neu Gene Expression in Osteosarcoma and Skeletal Ewing's Sarcoma Clin. Cancer Res., March 1, 2002; 8(3): 788 - 793. [Abstract] [Full Text] [PDF]
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A. Yuan, C.-J. Yu, K.-T. Luh, S.-H. Kuo, Y.-C. Lee, and P.-C. Yang Aberrant p53 Expression Correlates With Expression of Vascular Endothelial Growth Factor mRNA and Interleukin-8 mRNA and Neoangiogenesis in Non-Small-Cell Lung Cancer J. Clin. Oncol., February 15, 2002; 20(4): 900 - 910. [Abstract] [Full Text] [PDF]
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K. J. Savinainen, O. R. Saramaki, M. J. Linja, O. Bratt, T. L. J. Tammela, J. J. Isola, and T. Visakorpi Expression and Gene Copy Number Analysis of ERBB2 Oncogene in Prostate Cancer Am. J. Pathol., January 1, 2002; 160(1): 339 - 345. [Abstract] [Full Text] [PDF]
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I. Bieche, B. Parfait, S. Tozlu, R. Lidereau, and M. Vidaud Quantitation of androgen receptor gene expression in sporadic breast tumors by real-time RT-PCR: evidence that MYC is an AR-regulated gene Carcinogenesis, September 1, 2001; 22(9): 1521 - 1526. [Abstract] [Full Text] [PDF]
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I. Bièche, B. Parfait, V. Le Doussal, M. Olivi, M.-C. Rio, R. Lidereau, and M. Vidaud Identification of CGA as a Novel Estrogen Receptor-responsive Gene in Breast Cancer: An Outstanding Candidate Marker to Predict the Response to Endocrine Therapy Cancer Res., February 1, 2001; 61(4): 1652 - 1658. [Abstract] [Full Text]
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A. Yuan, C.-J. Yu, S.-H. Kuo, W.-J. Chen, F.-Y. Lin, K.-T. Luh, P.-C. Yang, and Y.-C. Lee Vascular Endothelial Growth Factor 189 mRNA Isoform Expression Specifically Correlates With Tumor Angiogenesis, Patient Survival, and Postoperative Relapse in Non-Small-Cell Lung Cancer J. Clin. Oncol., January 15, 2001; 19(2): 432 - 441. [Abstract] [Full Text] [PDF]
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M. Ferrero-Poüs, K. Hacène, C. Bouchet, V. Le Doussal, M. Tubiana-Hulin, and F. Spyratos Relationship between c-erbB-2 and Other Tumor Characteristics in Breast Cancer Prognosis Clin. Cancer Res., December 1, 2000; 6(12): 4745 - 4754. [Abstract] [Full Text]
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R. Lotan Are We Ready to Use Surrogate End Points and Surrogate Tissues to Evaluate Response to Chemopreventive and Therapeutic Intervention? Clin. Cancer Res., June 1, 2000; 6(6): 2126 - 2128. [Full Text]
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