Affiliations: [a] Department of Pathology, The Oncology Institute “Prof. Dr. Ion Chiricuta” Cluj-Napoca, Romania | [b] Department of Functional Genomics and Experimental Pathology, The Oncology Institute “Prof. Dr. Ion Chiricuta”, Cluj-Napoca, Romania | [c] Department of Immunology, University of Medicine and Pharmacy, “Iuliu Hatieganu”, Cluj-Napoca, Romania | [d] The Research Center for Functional Genomics, Biomedicine and Translational Medicine, “Iuliu Hatieganu” University of Medicine and Pharmacy, Cluj-Napoca, Romania | [x] The University of Medicine and Pharmacy Iuliu Hatieganu, Cluj, Napoca, Romania | [y] National Cancer Research Center, Istituto Tumori G Paolo II, IRCCS, Bari, Italy
Correspondence:
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Corresponding author: Ovidiu Balacescu, Department of Functional Genomics and Experimental Pathology, The Oncology Institute “Prof. Dr. Ion Chiricuta” Cluj-Napoca, 34-36 Republicii Str, cod 400015, Cluj-Napoca, Romania. Tel.: +40 264 590638; E-mail: [email protected].
Note: [1]
Authors with equal contribution to this work.
Abstract: Nowadays, HER2 testing in breast cancer represents a necessity for both prognostic and therapy. Despite widespread use of immunohistochemistry (IHC) for assessing HER2 status, there are some limitations to identify truly negative or positive HER2 cases. Fluorescence in situ hybridization (FISH) or chromogenic in situ hybridization (CISH) could solve the equivocal HER2 IHC cases but there is no consensus on which is the best method. Consequently, finding a sensitive method for HER2 testing is critical for the management of the disease. In addition, tumor heterogeneity is an important factor which could affect accuracy of molecular diagnostics. Laser capture micro-dissection (LCM) is used to isolate pure cell populations from heterogeneous tumor tissue. The combination between LCM and quantitative polymerase chain reaction (Q-PCR), the gold standard in molecular biology for quantifying gene amplification levels, could define an important tool to improve the molecular diagnostics of HER2 status. In our pilot study we used LCM and Q-PCR to evaluate HER2 gene amplification for invasive breast carcinoma samples. The samples were selected based on HER2 status assessed by IHC and CISH. Our results demonstrated high sensitivity of Q-PCR for assessing HER2 DNA amplification as well as a good concordance between Q-PCR and IHC/ CISH assay.
