title: Validation for histology-driven diagnosis in non-small cell lung cancer using hsa-miR-205 and hsa-miR-21 expression by two different normalization strategies.
abstract: Targeted therapy of non-small cell lung cancer (NSCLC) demands a more accurate tumor classification that is crucial for patient selection in personalized treatment. MicroRNAs constitute a promising class of biomarkers and a helpful tool for the distinction between lung adenocarcinoma (AC) and squamous cell lung carcinoma (SCC). The aim of this study was to evaluate the impact of two different normalization strategies, using U6 snRNA and hsa-miR-103 as reference genes, on hsa-miR-205 and hsa-miR-21 expression levels, in terms of the classification of subtypes of NSCLC. By means of a quantitative real-time polymerase chain reaction (qRT-PCR) microRNA expression levels were evaluated in a classification set of 98 surgically resected NSCLC fresh-frozen samples, and validated findings in an independent set of 42 NSCLC samples. The microRNA expression levels were exploited to develop a diagnostic test using two data normalization strategies. The performance of microRNA profiling in different normalization methods was compared. We revealed the microRNA-based qRT-PCR tests to be appropriate measures for distinguishing between AC and SCC (the concordance of histologic diagnoses and molecular methods greater than 88%). Performance evaluation of microRNA tests, based on the two normalization strategies, showed that the procedure using hsa-miR-103 as reference target has a slight advantage (sensitivity 83.33 and 100% in classification and validation set, respectively) compared to U6 snRNA. Molecular tests based on microRNA expression allow a reliable classification of subtypes for NSCLC and can constitute a useful diagnostic strategy in patient selection for targeted therapy.
authors: Department of Clinical Molecular Biology, Medical University of Bialystok, Bialystok, Poland.
authors: Radoslaw
authors: R
authors: Charkiewicz
authors: Department of Clinical Molecular Biology, Medical University of Bialystok, Bialystok, Poland.|Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
authors: Lothar
authors: L
authors: Pilz
authors: Department of Clinical Molecular Biology, Medical University of Bialystok, Bialystok, Poland.
authors: Anetta
authors: A
authors: Sulewska
authors: Department of Thoracic Surgery, Medical University of Bialystok, Bialystok, Poland.
authors: Miroslaw
authors: M
authors: Kozlowski
authors: Department of Histology and Embryology, Medical University of Bialystok, Bialystok, Poland.
authors: Wieslawa
authors: W
authors: Niklinska
authors: Department of Regenerative Medicine and Immune Regulation, Medical University of Bialystok, Bialystok, Poland.
authors: Marcin
authors: M
authors: Moniuszko
authors: Department of Medical Pathomorphology, Medical University of Bialystok, Bialystok, Poland.
authors: Joanna
authors: J
authors: Reszec
authors: Department of Clinical Molecular Biology, Medical University of Bialystok, Bialystok, Poland.|Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
authors: Christian
authors: C
authors: Manegold
authors: Department of Clinical Molecular Biology, Medical University of Bialystok, Bialystok, Poland.
authors: Jacek
authors: J
authors: Niklinski
mesh_terms: D000328:Adult; D000368:Aged; D002289:Carcinoma, Non-Small-Cell Lung / Q000175:diagnosis* / Q000235:genetics; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000235:genetics; D008297:Male; D035683:MicroRNAs / Q000032:analysis*; D008875:Middle Aged; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D035683:MicroRNAs; C534425:MIR205, human; C524940:MIRN21 microRNA, human
keywords: microRNA; non-small cell lung cancer; subclassification; target therapy