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Updated: May 12, 2026
Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay
Published on: August 14, 2018
Molecular determinations of EGFR and EML4-ALK on a single slide of NSCLC tissue
Paola Ulivi1, Maurizio Puccetti, Laura Capelli
1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST-IRCCS), Meldola, FC, Italy. p.ulivi@irst.emr.it
Introduction:
Tyrosine kinase inhibitors (TKIs) and anti-anaplastic lymphoma kinase (ALK) agents are highly effective for the treatment of non-small cell lung cancer (NSCLC) patients harbouring specific alterations, and molecular characterisation of the tumour is needed even when limited tumour material is available.
Methods:
20 patients with a known epidermal growth factor receptor (EGFR) gene status were enrolled: 10 had mutated and 10 had wild type tumours. FISH analysis was performed on one cytological or histological sample to determine EML4-ALK status, after which the same cells scraped off each slide were used to evaluate the EGFR status.
Results:
In the 10 EGFR mutated patients, molecular analysis showed the same results as those obtained before the FISH test. One patient with an EGFR mutation also showed an EML4-ALK translocation, and both FISH-positive and FISH-negative cells maintained the EGFR mutation.
Conclusions:
EGFR mutation analysis can be performed on the same sample previously submitted to the EML4-ALK FISH procedure.
Insights
Epidermal growth factor receptor (EGFR) mutation analysis is feasible on the same sample used for anaplastic lymphoma kinase (ALK) fluorescence in situ hybridization (FISH). This allows for comprehensive molecular characterization of non-small cell lung cancer (NSCLC) tumors.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Targeted therapies like tyrosine kinase inhibitors (TKIs) and anti-anaplastic lymphoma kinase (ALK) agents are crucial for treating non-small cell lung cancer (NSCLC).
- Accurate molecular characterization of tumors is essential for guiding treatment decisions, even with limited tissue samples.
Purpose of the Study:
- To determine if epidermal growth factor receptor (EGFR) mutation analysis can be performed on the same sample after anaplastic lymphoma kinase (ALK) fluorescence in situ hybridization (FISH).
- To assess the feasibility of sequential molecular testing on limited NSCLC tumor specimens.
Main Methods:
- Twenty NSCLC patients with known EGFR status were enrolled (10 mutated, 10 wild-type).
- Fluorescence in situ hybridization (FISH) was used to detect EML4-ALK status.
- The same cells from the FISH-tested slide were then used for EGFR mutation analysis.
Main Results:
- EGFR mutation analysis yielded consistent results when performed after FISH.
- In one patient with an EGFR mutation, an EML4-ALK translocation was also detected.
- EGFR mutations were maintained in both FISH-positive and FISH-negative cells.
Conclusions:
- EGFR mutation testing can be successfully performed on non-small cell lung cancer samples previously subjected to EML4-ALK FISH analysis.
- This approach supports comprehensive molecular profiling of NSCLC using limited tumor material.
