INVESTIGATION OF THE RELATIONSHIP OF TNFRSF11A GENE POLYMORPHISMS WITH BREAST CANCER DEVELOPMENT AND METASTASIS RISK IN PATIENTS WITH BRCA1 OR BRCA2 PATHOGENIC VARIANTS LIVING IN THE TRAKYA REGION OF TURKEY
Özdemir K, Gürkan H, Demir S, Atli E, Özen Y, Sezer A, Tunçbilek N, Çicin İ
*Corresponding Author: Hakan Gürkan, MD, PhD, Department of Medical Genetics, Genetic Diseases Diagnosis Center, Trakya University Faculty of Medicine, Balkan Campus, 22030 Edirne, Turkey. Tel: +90-533-218-8005. Fax: +90-284-235-7641. Email: dr_hakangurkan@yahoo.de, hgurkan@trakya.edu.tr
page: 49

MATERIALS AND METHODS

A total of 106 individuals were included in the study, of which 51 were patients diagnosed with breast cancer and 55 were healthy controls. The average age of those in the group diagnosed with breast cancer was 44.8, while the average age of those in the healthy control group was determined to be 44.0. Of the 51 breast cancer patients who were included in the patient group of the study, a total of 23 female patients had been living in the Trakya region of Turkey for at least three generations, were not related to each other, were diagnosed with breast cancer and were determined to have BRCA1 and/or BRCA2 mutations; 28 female patients presented without a BRCA1 and/or BRCA2 mutation. Of the 55 women in the control group, who volunteered to participate in the study, all were also living in the Trakya region for at least three generations, were not related to each other, were older than 18 years of age, were not diagnosed with familial breast cancer, had no history of breast-related disease during previous examinations and did not have any complaints about their breasts and ovaries. The Informed Volunteer Consent Form was signed by both the patients and the individuals in the control group, which contained information about the study and certified their approval. The study was approved by the Trakya University Faculty of Medicine Scientific Research Ethics Committee. The decision number was 16/03 with the numbered document TÜTF-BAEK 2016/219 on September 28, 2016 after the reason, purpose, approach and methods of the study were examined. Peripheral venous blood samples of 2.5 μL were drawn from the patients and those in the control groups, and genomic DNA was extracted using an EZ1 Advanced XL (Qiagen GmbH, Hilden, Germany) nucleic acid isolation device with the EZ1 DNA Blood 200 μL isolation kits (Qiagen GmbH). The concentration and purity values of the genomic DNA samples were measured with the Nano Drop 2000C device (Thermo Fisher Scientific, Wilmington, DE, USA) at wavelengths of 260/280 nm. Genomic DNA samples with an absorbance value at 1.5- 2.0 and a concentration value of 20.0-80.0 ng/μL were included in the study. Then, allelic discrimination was performed through a real-time polymerase chain reaction (qPCR) process using the TaqManŽ SNP Genotyping Assay kit (Applied Biosystems, Foster City, CA, USA) for rs4485469, rs9646629, rs34739845, rs17069904, rs884205 and rs4941129 SNPs. Statistical Analyses. In the study, patient and control group data were analyzed with the Statistical Package for the Social Sciences (SPSS) version 16.0 software (SPSS Inc., Chicago, IL, USA). Demographic data of the patient and control groups were interpreted by obtaining the numbers (n), %, χ2, mean, minimum and maximum values, and averages. The genotype frequencies of the rs4485469, rs9646629, rs34739845, rs17069904, rs884205 and rs4941129 SNPs in the TNFRSF11A gene of the breast cancer patients (with and without BRCA1 or BRCA2 mutations) and the control group were analyzed. In addition, TNFRSF11A rs4485469, rs9646629, rs34739845, rs17069904, rs884205, rs4941129 SNPs genotypes were compared according to metastasis, estrogen receptor (ER), progesterone receptor (PR), CerbB2 receptor positivity status of the patients with and without BRCA1/BRCA2 mutation.



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