P1. Molecular subclassification of NSCLC based on hsa-mir-205 and hsa-mir-21 expression using real time PCR
CELCC 2014 Abstracts

P1. Molecular subclassification of NSCLC based on hsa-mir-205 and hsa-mir-21 expression using real time PCR

Radoslaw Charkiewicz1, Miroslaw Kozlowski2, Anetta Sulewska1, Robert Milewski3, Wieslawa Niklinska4, Jacek Niklinski1,2

1Department of Clinical Molecular Biology, Medical University of Bialystok, Bialystok, Poland; 2Department of Thoracic Surgery, Medical University of Bialystok, Bialystok, Poland; 3Department of Statistic and Medical Informatics, Medical University of Bialystok, Bialystok, Poland; 4Department of Histology and Embryology, Medical University of Bialystok, Bialystok, Poland

Background: Routine histopathology is the current standard of lung cancer classification, but this time-honored method is unreliable. Nowadays, the new methods of treatment based on molecular targets emerge, and these approaches require using high precision in non-small cell lung cancer (NSCLCs) sub-classifications. MicroRNAs (miRNAs) are small, non-coding RNA molecules regulating gene expression by inducing mRNA degradation or by blocking translation. Recently, hsa-mir-205 was identified as a highly specific marker of stratified squamous epithelium with expression in squamous cell lung carcinoma. The objectives of our study were as follows: evaluation of expression levels of hsa-mir-205 and hsa-mir-21 on large group well-characterized, completely resected fresh frozen lung tumors, by LNA qRT- polymerase chain reaction (PCR), in order to predict histo-pathological subtype of NSCLC; an attempt to develop a molecular assay, based on the expression levels of hsa-mir-205 and hsa-mir-21 in tumor tissues, for precise discrimination of squamous cell carcinoma and adenocarcinoma of the lung.

Methods: Freshly frozen specimens of tumor tissues were obtained from resected 57 squamous and 41 adenocarcinoma of NSCLC. Levels of miRNA expression (hsa-mir-205, hsa-mir-21 and U6 snRNA as the endogenous control) were quantitated using the miRCURY LNA™ Universal RT microRNA PCR system (Exiqon). Sample score was then obtained using the formula Score according to Lebanony et al., which defines the normalized expression of hsa-mir-205 minus half the normalized expression of hsa-mir-21.

Results: We found a difference in the relative expression level of hsa-mir-205 (P<0.0001) and the “sample score” (P<0.0001) between the group of squamous and adenocarcinoma of NSCLC. The sensitivity and specificity of the developed molecular tests was 90.5% and 88.1% for the relative expression level of hsa-mir-205 and 87.5% and 89.3% for the “sample score”.

Conclusions: hsa-mir-205 is a highly useful marker for the differentiation of squamous cell carcinoma and adenocarcinoma of the lung. Molecular assay, based on the level of expression of hsa-mir-205 and the “sample score” can be used as potential tool in subclassification of NSCLC patients.

Keywords: Non-small cell lung cancer (NSCLC); tumor marker; tumor classification

doi: 10.3978/j.issn.2218-6751.2014.AB013

Cite this article as: Charkiewicz R, Kozlowski M, Sulewska A, Milewski R, Niklinska W, Niklinski N. Molecular subclassification of NSCLC based on hsa-mir-205 and hsa-mir-21 expression using real time PCR. Transl Lung Cancer Res 2014;3(5):AB013. doi: 10.3978/j.issn.2218-6751.2014.AB013

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