The Association Between rs11671784 Polymorphism in pre-miR-27a and Lung Cancer
AbstractLung cancer is the leading cause of cancer-related deaths worldwide and non-small cell lung cancer (NSCLC), especially adenocarcinoma, is the most common type in which the most important cases are related with KRAS or EGFR mutations or EGFR amplifications. MicroRNAs are the most remarkable controlling agents of gene expression. MiR-27a has two binding sites in 3'-untranslated-region of EGFR and three in KRAS. Moreover, it has a dual role in types of cancer and the expression decreases in some neoplasms, including the NSCLC. Hsa-mir-27a is located in the mir-23a~27a~24-2 cluster and the expression of these miRNAs is performed simultaneously but the down-regulation of miR-27a occurs independently. Polymorphisms in pre-miRNA can affect the miRNA processing and expression. In the current case-control study, we investigated the association between rs11671784 within the loop region of pre-miR-27a and lung cancer susceptibility. Genomic DNA was extracted from the blood samples of 110 healthy subjects and 70 patients using the salt procedure. After finishing the primer design, the genotype of rs11671784 was determined using the RFLP-PCR technique. A statistical analysis was performed on the Power Marker, SPSS version-23 software and the SISA website.The present study examines the correlation between rs11671784 and lung cancer for the first time, which proved that the presence of the T allele at the polymorphism position can reduce the risk of lung cancer up to 6.7 times (OR=0.15, p=0.039), likely because of influencing the processing of miR-27a. These results suggest that rs11671784 could be used as a resistance marker for the genetic susceptibility to lung cancer among the Isfahan population.
Zagryazhskaya A, Zhivotovsky B. miRNAs in lung cancer: a link to aging. Ageing Res Rev 2014; 17: 54-67.
Horn, L; Pao W; Johnson DH. Chapter 89. In: Longo DL, Kasper DL, Jameson JL, et al. Harrison's principles of internal medicine . McGraw-Hill. 2012; 18.
Lu C, Onn A, Vaporciyan AA, et al. Cancer of the lung. Holland-Frei Cancer Medicine. People's Medical Publishing House 2010; 8.
Larsen JE, Minna JD. Molecular biology of lung cancer: clinical implications. Clin Chest Med 2011; 32: 703-740.
Babashah S, Sadeghizadeh M, Tavirani MR, Farivar S and Soleimani M. Aberrant microRNA expression and its implications in the pathogenesis of leukemias. Cellular Oncology 2012; 355: 317-334.
Kasahara Y, Nakamura RM and Kim PS. The role of micro-RNAs in cancer. Grody WW, Nakamura RM, Kiechle FL, Strom Ch. Techniques and Applications for the Clinical Laboratory. Molecular Diagnostics 2009; 1: 205-14.
Chen H, Goldberg MS, Villeneuve PJ. A systematic review of the relation between long-term exposure to ambient air pollution and chronic diseases. Rev Environ Health 2008; 23: 243-297.
The UCSC Genome Browser, the University of California, Santa Cruz, the United States: http://genome.ucsc.edu Accessed 
Roskoski R, Jr. The ErbB/HER family of protein-tyrosine kinases and cancer. Pharmacol Res 2014; 79: 34-74.
Chen KY, Hsiao CF, Chang GC, Tsai YH, Su WC, Chen YM, et al. "EGFR polymorphisms, hormone replacement therapy and lung adenocarcinoma risk: analysis from a genome-wide association study in never-smoking women." Carcinogenesis 2013; 34(3): 612-619.
Yuan Z, Shin J, Wilson A, Goel S, Ling YH, Ahmed N, et al. An A13 repeat within the 3'-untranslated region of epidermal growth factor receptor (EGFR) is frequently mutated in microsatellite instability colon cancers and is associated with increased EGFR expression. Cancer Res 2009; 69: 7811-7818.
Center for Computational Research SUNY at Buffalo, The State University of New York: http://mirdsnp.ccr.buffalo.edu/gene_display?id=4d88de4a7a87940f06002ce6&a=tscan&hd=0 Accessed 
Chhabra R, Dubey R, Saini N. Cooperative and individualistic functions of the microRNAs in the miR-23a~27a~24-2 cluster and its implication in human diseases. Mol Cancer 2010; 9: 232.
Zhang Z, Liu S, Shi R, Zhao G. miR-27 promotes human gastric cancer cell metastasis by inducing epithelial-to-mesenchymal transition. Cancer Genet 2011; 204: 486-491.
MicroRNA-Target Interactions, National Chiao Tung University, the United States: http://mirtarbase.mbc.nctu.edu.tw Accessed 
Wang N, Lu H, Chen W, Gan M, Cao X, Zhang J, Chen L. Primary microcephaly gene MCPH1 shows a novel molecular biomarker of human renal carcinoma and is regulated by miR-27a. Int J Clin Exp Pathol 2014; 7: 4895-4903.
Acunzo M, Romano G, Palmieri D, Lagana A, Garofalo M, Balatti V, et al. Cross-talk between MET and EGFR in non-small cell lung cancer involves miR-27a and Sprouty2. Proc Natl Acad Sci U S A 2013; 110: 8573-8578.
Heegaard NHH, Schetter AJ, Welsh JA, Yoneda M, Bowman ED, Harris CC. Circulating micro‐RNA expression profiles in early stage nonsmall cell lung cancer. Int J Cancer 2012; 130(6): 1378-1386
Wang Q, et al. Upregulation of miR-27a contributes to the malignant transformation of human bronchial epithelial cells induced by SV40 small T antigen. Oncogene 2011; 30: 3875-3886.
Mujahid S, Logvinenko T, Volpe MV, Nielsen HC. miRNA regulated pathways in late stage murine lung development. BMC Dev Biol 2013; 13: 13.
Xiong X, Kang X, Zheng Y, Yue S, Zhu S. Identification of loop nucleotide polymorphisms affecting microRNA processing and function. Mol Cells 2013; 36: 518-526.
Xu J, Tian S, Yin Z, et al. MicroRNA-binding site SNPs in deregulated genes are associated with clinical outcome of non-small cell lung cancer. Lung Cancer 2014; 85(3): 442-448.
Miller SA, Dykes DD and Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Research 1988; 16: 1215.
National Center for Biotechnology Information, dbSNP VCF Files, the United States: https://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?rs=11671784 Accessed 
The first health information database, Iran: http://www.salamatnews.com/news/25155 Accessed 
Song B, Yan G, Hao H, Yang B. rs11671784 G/A and rs895819 A/G polymorphisms inversely affect gastric cancer susceptibility and miR-27a expression in a Chinese population. Med Sci Monit 2014; 20: 2318-2326.
Deng Y, Bai H, Hu H. rs11671784 G/A variation in miR-27a decreases chemo-sensitivity of bladder cancer by decreasing miR-27a and increasing the target RUNX-1 expression. Biochem Biophys Res Commun 2015; 458: 321-327.
Salzman DW, Weidhaas JB. SNPing cancer in the bud: microRNA and microRNA-target site polymorphisms as diagnostic and prognostic biomarkers in cancer. Pharmacol Ther 2013; 137: 55-63.
Shen J, Xia W,Khotskaya Y, et al. EGFR modulates microRNA maturation in response to hypoxia through phosphorylation of AGO2. Nature 2013; 497: 383-387.
MicroRNA SNP Database: http://www.bioguo.org/miRNASNP Accessed 
The RNAfold web server, Institute for Theoretical Chemisty, University of Vienna: http://rna.tbi.univie.ac.at/cgi-bin/RNAfold.cgi Accessed 
Tafsiri E, Javadi A, Karimipoor M. Circulating microRNAs in serum and other body fluids as diagnostic of cancer. Genetics in Third Millennium 2014; 2: 3472-3587.
Lim LP, Glasner ME, Yekta S, Burge CB, Bartel DP. Vertebrate microRNA genes. Science 2003; 299: 1540.
Cho WC. MicroRNAs: potential biomarkers for cancer diagnosis, prognosis and targets for therapy. Int J Biochem Cell Biol 2010; 42: 1273-1281.
Ruan K, Fang X, Ouyang G. MicroRNAs: novel regulators in the hallmarks of human cancer. Cancer Lett 2009; 285: 116-126.
Negrini M, Nicoloso MS and Calin GA. MicroRNAs and cancer new paradigms in molecular oncology. Current opinion in cell biology 2009; 21: 470-479
Yang R, Burwinkel B. A bias in genotyping the miR-27a rs895819 and rs 11671784 variants. Breast Cancer Res Treat 2012; 134: 899-901.