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Updated: May 15, 2026

Ultra-Fast Amplicon-Based Next-Generation Sequencing in Non-Squamous Non-Small Cell Lung Cancer
Published on: September 8, 2023
Molecular profiling of thin-prep FNA samples in assisting clinical management of non-small-cell lung cancer
Daniela Petriella1, Domenico Galetta, Vincenza Rubini
1National Cancer Research Centre, Istituto Tumori "Giovanni Paolo II", v. Orazio Flacco 65, 70124 Bari, Italy.
Abstract:
The discovery of new target treatments for NSCLC has led to a search for new genetic and epigenetic markers able to selectively predict response to these new drugs. Somatic mutations in EGFR and KRAS genes are routinely analyzed to predict response to tyrosine kinase inhibitors (TKIs), used in the treatment of NSCLC patients, whose efficacy depend on the presence or the absence of specific mutations. MicroRNA (miRNA) expression evaluation has been recently analyzed because of the involvement of these molecules in lung cancer pathogenesis and in drug resistance. Only 30 % of NSCLC patients present a resectable stage at time of diagnosis so tissue samples cannot be the only starting material for genetic and epigenetic analysis. Therefore, the possibility to use cytological sampling already used for diagnosis also for molecular testing is emerging. The aim of this study was to evaluate for the first time in lung cancer the use of liquid-based cytology both for EGFR and KRAS mutational testing and for the expression trend of some miRNAs involved in lung cancer pathogenesis: miR-21, miR-155, miR-7, and let7a. We enrolled 20 fine-needle aspirate (FNA) samples diagnosed as NSCLC, 10 FNAs without neoplastic cells, and tissue samples coming from 5 of the 20 patients who underwent surgery after FNA NSCLC diagnosis. All Thin-Prep processed FNA samples were evaluable for DNA and RNA analysis and results were compared with those of the small group of patients whose matched tumor histology was available. The mutational status of the EGFR and KRAS genes and the expression profile of the selected miRNA showed comparable results between FNA samples and histological tissues. Our results underline that cytological samples could give the same genetic information as that obtained from histological specimens and so could be collected to create a nucleic acids bank.
Insights
Liquid-based cytology effectively analyzes EGFR and KRAS mutations and microRNA expression in non-small cell lung cancer (NSCLC). This approach provides genetic information comparable to tissue samples, enabling molecular testing from cytological specimens.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Targeted therapies for non-small cell lung cancer (NSCLC) require predictive genetic and epigenetic markers.
- EGFR and KRAS mutations are key for predicting response to tyrosine kinase inhibitors (TKIs).
- MicroRNAs (miRNAs) play a role in lung cancer pathogenesis and drug resistance.
Purpose of the Study:
- To evaluate liquid-based cytology for EGFR/KRAS mutational testing and miRNA expression in NSCLC.
- To compare molecular analysis results from cytological samples with histological tissues.
- To explore the potential of cytological samples for nucleic acid banking.
Main Methods:
- Analysis of 20 NSCLC fine-needle aspirate (FNA) samples using liquid-based cytology (Thin-Prep).
- EGFR and KRAS mutational testing and expression analysis of miR-21, miR-155, miR-7, and let7a.
- Comparison of results with matched surgical tissue samples from 5 patients.
Main Results:
- Liquid-based cytology samples were evaluable for both DNA and RNA analysis.
- Comparable EGFR/KRAS mutational status and miRNA expression profiles were observed between FNA samples and histological tissues.
- Cytological samples yielded consistent genetic information compared to tissue specimens.
Conclusions:
- Liquid-based cytology is a viable method for molecular testing in NSCLC.
- Cytological samples can serve as a reliable source for genetic and epigenetic analysis in NSCLC.
- This technique facilitates the creation of nucleic acid banks from routinely collected cytological specimens.
