Balkan Journal of Medical Genetics

MITOCHONDRIAL DNA 4977 bp DELETION IN CHRONIC CERVICITIS AND CERVIX CANCERS
Kara M, Tatar A, Borekci B, Dagli F, Oztas S
*Corresponding Author: Murat Kara M.D., Department of Genetics, School of Medicine, Firat University, Universite cad. No. 2, 23119 Elazig, Turkey; Tel: +90-424-233-35-55/1938; Fax: +90-424-238-80-96; E-mail: drmuratkara@hotmail.com
page: 25

MATERIAL AND METHODS

Samples and DNA Isolation. This study was conducted during the years 2008-2009 at Ataturk University Faculty of Medicine, Department of Medical Genetics, Department of Pathology, Department of Obstetrics and Gynecology, Erzurum, Turkey, and Firat University Faculty of Medicine Department of Pathology, Elazig, Turkey. The study included 72 participants in an East Turkish population; 35 patients with chronic cervicitis, 21 patients with cervical cancer, and 16 samples taken from formalin-fi xed, paraffi n-embedded normal cervix tissues (FFPTs). Ethical approval was obtained by the Ataturk University Ethics Committee for the collection and analysis of the samples. The mean ages of the patients with chronic cervicitis was 50.2 ± 8.9 (40-72 years), of the patients with cervix cancer it was 62.4 ± 9.9 (47-79 years) and of the samples from the control group it was 45.5 ± 8.6 (28-61 years). Primers, DNA Extraction and Polymerase Chain Reaction Amplifi cation. In order to demonstrate the 4977 bp deletion of mtDNA, the preparation of primers based on gene sequences was obtained from the database website http://www. mitomap.org/. Nucleotide sequences of purchased primers (Bio Basics Inc., Markham, ON, Canada) are shown in the Table 1. These primer pairs contained two control regions as negative and positive controls and one deletion region. DNA extraction was performed on 5 mg fresh tissue samples of individuals diagnosed as having chronic cervicitis and cervical malignancy by using the DNA isolation kit (GENTRA-Genomic DNA Purifi cation Kit; Madison, WI, USA). For the control group, samples taken from FFPTs were used for DNA extraction by using the DNA isolation kit (DZDNA-Paraffi n Embedded Tissue DNA Isolation Kit, Dr. Zeydanli, Ankara, Turkey). The PCR reactions (50 μL) were performed in 10 μM Tris-HCl, pH 8.4, 50 μM KCl, 2 μM MgCl2, 200 μM each of dNTP, 20 pmol of each primer and 5 U Taq polymerase (Fermentas Inc., Hanover, MD, USA) in a thermocycler under the following conditions: 95°C for 1 min. (initial denaturation) followed by 31 cycles at 94°C for 25 seconds (denaturation), 60°C for 35 seconds (annealing), 72°C for 1 min. (extension), and a fi nal extension at 72°C for 10 min. An aliquot (10 μL) of each amplicon was examined on 1.8% agarose gels and stained with ethidium bromide. During visualization, a 470 bp fragment for the deletion region, a 1029 bp fragment for the positive control site outside the deletion region and a 564 bp fragment for the negative control site in the deletion region were observed. Statistical Analyses. All statistical analyses were performed by using the SPSS for Windows computing program version 12.0. Disease type and mtDNA results were tested by chi-square. A p value lower than 0.05 was accepted as statistically significant.

Number 27 VOL. 27 (2), 2024	Number 27 VOL. 27 (1), 2024
Number 26 Number 26 VOL. 26(2), 2023 All in one	Number 26 VOL. 26(2), 2023
Number 26 VOL. 26, 2023 Supplement	Number 26 VOL. 26(1), 2023
Number 25 VOL. 25(2), 2022	Number 25 VOL. 25 (1), 2022
Number 24 VOL. 24(2), 2021	Number 24 VOL. 24(1), 2021
Number 23 VOL. 23(2), 2020	Number 22 VOL. 22(2), 2019
Number 22 VOL. 22(1), 2019	Number 22 VOL. 22, 2019 Supplement
Number 21 VOL. 21(2), 2018	Number 21 VOL. 21 (1), 2018
Number 21 VOL. 21, 2018 Supplement	Number 20 VOL. 20 (2), 2017
Number 20 VOL. 20 (1), 2017	Number 19 VOL. 19 (2), 2016
Number 19 VOL. 19 (1), 2016	Number 18 VOL. 18 (2), 2015
Number 18 VOL. 18 (1), 2015	Number 17 VOL. 17 (2), 2014
Number 17 VOL. 17 (1), 2014	Number 16 VOL. 16 (2), 2013
Number 16 VOL. 16 (1), 2013	Number 15 VOL. 15 (2), 2012
Number 15 VOL. 15, 2012 Supplement	Number 15 Vol. 15 (1), 2012
Number 14 14 - Vol. 14 (2), 2011	Number 14 The 9th Balkan Congress of Medical Genetics
Number 14 14 - Vol. 14 (1), 2011	Number 13 Vol. 13 (2), 2010
Number 13 Vol.13 (1), 2010	Number 12 Vol.12 (2), 2009
Number 12 Vol.12 (1), 2009	Number 11 Vol.11 (2),2008
Number 11 Vol.11 (1),2008	Number 10 Vol.10 (2), 2007
Number 10 10 (1),2007	Number 9 1&2, 2006
Number 9 3&4, 2006	Number 8 1&2, 2005
Number 8 3&4, 2004	Number 7 1&2, 2004
Number 6 3&4, 2003	Number 6 1&2, 2003
Number 5 3&4, 2002	Number 5 1&2, 2002
Number 4 Vol.3 (4), 2000	Number 4 Vol.2 (4), 1999
Number 4 Vol.1 (4), 1998	Number 4 3&4, 2001
Number 4 1&2, 2001	Number 3 Vol.3 (3), 2000
Number 3 Vol.2 (3), 1999	Number 3 Vol.1 (3), 1998
Number 2 Vol.3(2), 2000	Number 2 Vol.1 (2), 1998
Number 2 Vol.2 (2), 1999	Number 1 Vol.3 (1), 2000
Number 1 Vol.2 (1), 1999	Number 1 Vol.1 (1), 1998

About the journal ::: Editorial ::: Subscription ::: Information for authors ::: Contact