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Validation of a differential PCR and an ELISA procedure in studying HER ‐2/ neu status in breast cancer
Author(s) -
Valerón Pilar F.,
Chirino Ricardo,
Fernandez Leandro,
Torres Santiago,
Navarro Domingo,
Aguiar José,
Cabrera Juan J.,
DiazChico Bonifacio N.,
DiazChico Juan C.
Publication year - 1996
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19960117)65:2<129::aid-ijc1>3.0.co;2-0
Subject(s) - breast cancer , oncogene , gene , cancer , gene expression , microbiology and biotechnology , biology , differential diagnosis , cancer research , pathology , medicine , genetics , cell cycle
HER ‐2/ neu oncogene status and total cellular p185 HER‐2 content were simultaneously analyzed in 415 invasive breast‐cancer specimens by differential PCR and ELISA respectively. Mathematical analysis of the data led us to establish a cut‐off value of 1.7 for the ratio between the intensity of the HER ‐2/ neu gene band and the reference gene band, to consider the HER ‐2/ neu gene amplified, and of 260 fmol/mg protein, to consider p185 HER‐2 over‐expressed. Of the 415 tumors studied, 15% showed a diverse degree of HER ‐2/ neu gene amplification. Of these tumors, 87% showed over‐expression of the p185 HER‐2 . Of the remaining 352 specimens that did not display HER ‐2/ neu gene amplification, 97% showed no p185 HER‐2 over‐expression ( p < 0.0001). In 40 selected samples with a p185 HER‐2 level lower than 260 fmol/mg protein, the degree of p185 HER‐2 phosphorylation was very low or undetectable. Conversely, 38 of 46 selected tumors with a p185 HER‐2 level higher than 260 fmol/mg protein exhibited a considerable degree of p185 HER‐2 phosphorylation ( p < 0.0001). Our data suggest that: (i) differential PCR and ELISA, which are relatively simple procedures, give similar information on HER ‐2/ neu status in breast cancer; and (ii) given the large series analyzed, the cutoff values established can be considered as safe values for determining whether, in a given tumor, the HER ‐2/ neu oncogene is amplified or p185 HER‐2 is over‐expressed. © 1996 Wiley‐Liss, Inc.