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Clinical Chemistry 43: 2114-2120, 1997;
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(Clinical Chemistry. 1997;43:2114-2120.)
© 1997 American Association for Clinical Chemistry, Inc.

Articles

Quantification of c-erbB-2 gene expression in breast cancer by competitive RT-PCR

Françoise Révillion, Louis Hornez and Jean-Philippe Peyrat2
We adapted competitive reverse transcription-polymerase chain reaction (RT-PCR) for quantitative evaluation of c-erbB-2 expression in breast cancer. A fixed amount of cDNA target was coamplified with dilutions of a nonhomologous DNA sequence used as an internal standard (IS). The IS and the target shared the same primer sequences but yielded PCR products of different sizes (348 and 340 bp, respectively). A fluorescent sense primer was used so that the PCR products separated on denaturing polyacrylamide gels could be quantified with an automated DNA sequencer. In human breast cancer cell lines, c-erbB-2 expression was found to range from 0.151 to 652 zmol/µg total RNA (i.e., 91 to 391 200 molecules/µg total RNA; 1 zmol = 10-3 amol), with the two highest values corresponding to the c-erbB-2 overexpressing cell lines MDA-MB-453 and SK-BR-3. In a series of 39 breast cancer biopsies, the concentrations ranged from 0.117 zmol/µg to 1.15 amol/µg total RNA (i.e., 70 to 690 000 molecules/µg total RNA). The c-erbB-2 oncoprotein (p185) was determined in 30 samples by an enzyme immunoassay. A close correlation was found between c-erbB-2 gene and oncoprotein expression (P = 0.0067). Thus, this competitive RT-PCR method appears to be a reliable way to evaluate the expression of c-erbB-2 in small tumor samples.




The following articles in journals at HighWire Press have cited this article:


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F. Revillion, V. Lhotellier, L. Hornez, J. Bonneterre, and J.-P. Peyrat
ErbB/HER ligands in human breast cancer, and relationships with their receptors, the bio-pathological features and prognosis
Ann. Onc., January 1, 2008; 19(1): 73 - 80.
[Abstract] [Full Text] [PDF]

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H. Xie, Y. H. Yu, P.-L. Mao, and Z. Gao
Highly sensitive amperometric detection of genomic DNA in animal tissues
Nucleic Acids Res., January 20, 2004; 32(2): e15 - e15.
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M. Konigshoff, J. Wilhelm, R. M. Bohle, A. Pingoud, and M. Hahn
HER-2/neu Gene Copy Number Quantified by Real-Time PCR: Comparison of Gene Amplification, Heterozygosity, and Immunohistochemical Status in Breast Cancer Tissue
Clin. Chem., February 1, 2003; 49(2): 219 - 229.
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G. Castaldo, G. Calcagno, R. Sibillo, R. Cuomo, G. Nardone, L. Castellano, C. Del Vecchio Blanco, G. Budillon, and F. Salvatore
Quantitative Analysis of Aldolase A mRNA in Liver Discriminates between Hepatocellular Carcinoma and Cirrhosis
Clin. Chem., July 1, 2000; 46(7): 901 - 906.
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I. Bieche, P. Onody, I. Laurendeau, M. Olivi, D. Vidaud, R. Lidereau, and M. Vidaud
Real-Time Reverse Transcription-PCR Assay for Future Management of ERBB2-based Clinical Applications
Clin. Chem., August 1, 1999; 45(8): 1148 - 1156.
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Copyright © 1997 by the American Association for Clinical Chemistry.