FISH molecular testing in cytological preparations from solid tumors
© Caria and Vanni; licensee BioMed Central Ltd. 2014
Received: 12 May 2014
Accepted: 30 July 2014
Published: 22 August 2014
Many of the exciting new developments in solid tumor molecular cytogenetics impact classical and molecular pathology. Fluorescence in situ hybridization to identify specific DNA target sequences in nuclei of non-dividing cells in solid neoplasms has contributed to the integration of molecular cytogenetics into cytology in spite of the remarkable promiscuity of cancer genes. Indeed, although it is a low-throughput assay, fluorescence in situ hybridization enables the direct disclosure and localization of genetic markers in single nuclei. Gene fusions are among the most prominent genetic alterations in cancer, providing markers that may be determinant in needle biopsies that are negative or suspicious for malignancy, and may contribute to the correct classification of the tumors. In view of the expanding use of fluorescence in situ hybridization in cytology, future challenges include automated sample evaluation and the specification of common criteria for interpreting and reporting results.
KeywordsFISH Gene fusions Cytology Solid tumor Gene promiscuity
Various types of genetic alterations, as well as epigenetic phenomena, have been identified and are now considered important in the classification, prognosis, and treatment of cancer. Correlations between genomic instability and carcinogenesis have been extensively investigated, leading to the recognition of an increasing number of genetic abnormalities as a tumor driving force. Currently, several molecular approaches are available to investigate tumor cell pathobiology at different levels (chromosome, gene, gene expression). The predominant approaches include immunohistochemistry, fluorescence in situ hybridization (FISH), polymerase chain reaction, array-based and omics-based techniques –. The integration of results obtained using these platforms has been invaluable in clarifying genetic alterations associated with cancer and in interpreting the key role of the impaired signaling pathways. Gene gains and losses and gene disruptions by chromosome translocation, inversion, or deletion have been recognized as playing a pathogenetic role in many cancers. These exciting new developments in solid tumor molecular cytogenetics impact classical and molecular pathology, and an increasing number of chromosome markers have been integrated into World Health Organization tumor classifications . Some of these markers are also relevant to selection of therapies targeting the protein products of gene fusions. In this scenario the impact of testing gene alterations by interphase FISH in material from needle biopsies and organic fluids has rapidly increased.
Promiscuity: a false dilemma?
Cytology and cytogenetics
Considering the introduction of systematic genomic testing for some tumors (such as lung and breast cancer) ,, the consequent need for a correct evaluation of ratio value in the presence of genetic heterogeneity , and the growing demand for FISH tests in fine needle aspirations and organic fluids, two main challenges for the future can be foreseen: the implementation of automated FISH evaluation and the specification of common criteria for interpreting and reporting FISH results in as many tumor types as possible. A significant impediment to evaluating the ever increasing numbers of clinical FISH tests requested is imposed by the labor intensive nature of the assay, as each test requires scoring numerous interphase nuclei by double blind observation. Automated FISH, with strictly established parameters for standardization, could partly overcome these issues, although automation has yet to be perfected . Specific recommendations and guidelines for FISH on tumors have been established within ACMG (American College of Medical Genetics and Genomics)  and E.C.A (European Cytogeneticists Association) . On the other hand, common objective interpretation criteria for FISH on cytological preparations, as well as quality control and quality assurance policies, remain limited ,, and require an extraordinary cooperative effort and interaction between cytogeneticists and cytologists. It would be desirable to convene expert advisory panels from scientific societies of clinical cytogeneticists and pathologists to establish evaluation criteria for the various tumors, based on expertise and a review of published literature, with a view to establishing common shared recommendations.
Many of the exciting new developments of molecular cytogenetics are having a profound impact on classical and molecular cytology. The growing demand for cytological FISH tests highlights the need for the specification of common criteria for interpreting and reporting FISH results, for quality control and quality assurance policies, and for possible implementation of automated FISH evaluation.
PC and RV participated in commentary design and wrote the manuscript. They read and approved the final manuscript.
We wish to express our gratitude to Daniela V Frau for providing us with Figure 4. Manuscript preparation was funded by Fondazione Banco di Sardegna, grant prot No 17741 and POR Sardegna-FSE 2007–2013.
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