Assessment of association between genetic variants in microRNA genes hsa-miR-499, hsa-miR-196a2 and hsa-miR-27a and prostate cancer risk in Serbian population Z. Nikolić a , D. Savić Pavićević a , N. Vučić a , S. Cidilko a , N. Filipović b , S. Cerović c , V. Vukotić b , S. Romac a , G. Brajušković a, ⁎ a University of Belgrade, Faculty of Biology, Belgrade, Serbia b Department of Urology, Clinical Centre “dr Dragiša Mišović”, Belgrade, Serbia c Institute of Pathology, Military Medical Academy, Belgrade, Serbia abstract article info Article history: Received 2 June 2015 Accepted 18 June 2015 Available online 22 June 2015 Keywords: Prostate cancer MicroRNA Association study miR-499 miR-196a2 miR-27a Due to their potentially functional significance, genetic variants within microRNA genes have been recognized as candidates for cancer-related genetic biomarkers. Among the most extensively studied so far are rs3746444, rs11614913 and rs895819. Nevertheless, only few previous studies in Asian population analyzed the association of rs3746444 and rs11614913 with prostate cancer (PCa) risk, while rs895819 was not evaluated in relation to this issue. The aim of this study was to assess the possible association between these genetic variants and PCa risk and progression in Serbian population. 355 samples of peripheral blood were obtained from the patients with PCa and 353 samples from patients with benign prostatic hyperplasia (BPH). 312 volunteers derived from general population who gave samples of buccal swabs were included in the control group. Genotyping of rs3746444, rs11614913 and rs895819 was performed by using PCR-RFLP method, HRM analysis and allele- specific PCR, respectively. Allelic and genotypic associations were evaluated by unconditional linear (for serum PSA level in PCa patients) and logistic regression method with adjustment for age. Minor allele C of rs895819 was found to be associated with the increased risk of developing PCa under dominant (P = 0.035; OR = 1.38, 95%CI 1.02–1.86) and overdominant (P = 0.04; OR = 1.37, 95%CI 1.01–1.85) genetic model. Same genetic variant was found to be associated with the clinical stage of localized PCa, as well as with the presence of distant metastases. Allele G of rs3746444 was also shown to be associated with the decreased risk of PCa progression. According to our data, rs3746444 qualifies for a genetic variant potentially associated with PCa aggressiveness in Serbian population. Furthermore, our study provided the first evidence of association between rs895819 and PCa risk, as well as for its genetic association with the presence of distant metastases among PCa patients. © 2015 Elsevier Inc. All rights reserved. 1. Introduction According to recent projections based on GLOBOCAN 2012 report, global cancer burden is expected to substantially increase within the next decade (Ferlay et al., 2015). Being the second leading cancer in terms of incidence and ranking sixth among causes of cancer-related death worldwide, prostate cancer (PCa) is due to significantly contrib- ute to this growing public health problem (Bray et al., 2012; Ferlay et al., 2015). Among diagnosed tumors, clinically inapparent latent prostate adenocarcinomas which do not require aggressive therapy, but active surveillance, represent a significant proportion (Van der Kwast and Roobol, 2013). Therefore, identification of genetic variants contributing not only to cancer susceptibility, but also to PCa aggressive- ness has become one of the imperatives in research activity regarding prostate carcinogenesis. These genetic variants are considered to be po- tential nonstandard prognostic parameters of PCa that could be imple- mented in algorithms used for assessing the risk of PCa development, as well as for evaluating the risk of PCa progression and outcome predic- tion (Goh and Eeles, 2014; Helfand and Catalona, 2014). Although large genome-wide association studies (GWAS) have made the major contribution to identifying genetic factors contributing to prostate cancer susceptibility, candidate-gene based approaches have also revealed multiple PCa associated regions (Goh and Eeles, 2014). Due to accumulating evidence of the involvement of regulatory mechanisms based on the activity of non-coding RNAs in prostate carcinogenesis, Experimental and Molecular Pathology 99 (2015) 145–150 Abbreviations: PCa, prostate cancer; BPH, benign prostatic hyperplasia; GWAS, genome-wide association study; PSA, prostate-specific antigen; GS, Gleason score; PCR– RFLP, restriction fragment length polymorphism analysis of PCR-amplified fragments; HRMA, high resolution melting analysis; HWE, Hardy–Weinberg equilibrium; OR, odds ratio; CI, confidence interval; AIC, Akaike information criterion. ⁎ Corresponding author. E-mail address: brajuskovic@bio.bg.ac.rs (G. Brajušković). http://dx.doi.org/10.1016/j.yexmp.2015.06.009 0014-4800/© 2015 Elsevier Inc. All rights reserved. Contents lists available at ScienceDirect Experimental and Molecular Pathology journal homepage: www.elsevier.com/locate/yexmp