Molecular diagnosis in metastatic cancer. molise experience

Abstract

Background: The Molecular Oncogenetic Laboratory's arises to support medical therapy for metastatic cancer and to benefit cancer patients, come from the Oncology Units of Molise Hospitals, of “target therapy” for the treatment of colorectal, lung, breast and gastric cancers, melanoma and CML. Materials/methods: The laboratory has two sections: FISH and molecular. By FISH we assess the status of HER2 in breast and gastric cancer with Leica BOND system HER2 FISH probes LSI® HER2/CEP17® FISH PathVysion®. The processing of samples is standardized in a semi-automatic system and the interpretation of fluorescent signals is done according to the reccomendations Asco/Cap 2013. By FISH, in manual mode, we evaluate the translocation BCR/ABL in CML. Real Time-PCR, by kits EntroGene®, is used to assess the status of all-RAS, EGFR and BRAF genes respectively in colorectal carcinoma, lung adenocarcinoma and melanoma. RT-PCR involve the use of mutation-specific probes, that due to cancer genetics heterogeneity, are not always able to detect less frequent mutations. The cyto-histological samples used are evaluated for cellularity and suitability analysis. The protocols used comply with the recommendations AIOM/SIAPEC-IAP. Results: From 2014 to date we have been analyzed 116 samples. 46 subjected to FISH (41 breast and 5 stomach). In 32 cases FISH resulted negative, in 9 positive and 5 samples were unsuitable for analysis. 6 samples were tested for diagnosis or follow-up of LMC. In 3 was present t (9; 22). In molecular biology, we evaluated 64 samples (12 EGFR, 13 BRAF, 39 RAS). 43 samples came from primary cancer and 21 from metastases. The results were: 2 mutated (ex19del) and 10 wt for EGFR; 6 mutated (V600E) and 7 wt for BRAF; 22 mutated (9 for G12D, 3 for G12V and G13D, 2 for G12a and A146X, 1 for Q61R, G13C and G13V) and 16 wt for KRAS; 1 NRAS mutated (Q61R). Conclusion: As soon as possible we will expand the panel of oncogenes with FISH probes (MM, LLA, LLC, ALK rearrangements). It is hoped that the Laboratory will become a reference center for the entire region Molise. The laboratory adheres to programs of National Quality Control for the certification of scientific procedures and diagnostic data products.