Association of serum paraoxonase 1 activities, polymorphisms and oxidative stress in breast cancer patients with type 2 diabetes mellitus

J Med Biochem 2019; 38 (3) DOI: 10.2478/jomb-2018-0043 UDK 577.1 : 61 ISSN 1452-8258 J Med Biochem 38: 368 –375, 2019 Original paper Originalni nau~ni rad ASSOCIATION OF SERUM PARAOXONASE 1 ACTIVITIES, POLYMORPHISMS AND OXIDATIVE STRESS IN BREAST CANCER PATIENTS WITH TYPE 2 DIABETES MELLITUS VEZA AKTIVNOSTI SERUMSKE PARAOKSONAZE 1, POLIMORFIZMA I OKSIDATIVNOG STRESA KOD PACIJENTKINJA SA RAKOM DOJKE I DIJABETESOM TIP 2 Fatma Ceyla Eraldemir1, Nihal Üren2, Tuğba Kum1, Burcu Erbay3, Deniz Şahin4, Emel Ergül2, Esra Acar1, Doğa Özsoy1, Mustafa Çekmen5, Hale Kır1, Zafer Utkan3 1 2 Department of Biochemistry, School of Medicine, Kocaeli University, Kocaeli, Turkey Department of Medical Biology and Genetics, School of Medicine, Kocaeli University, Kocaeli, Turkey 3 Department of General Surgery, School of Medicine, Kocaeli University, Kocaeli, Turkey 4 Department of Physiology, School of Medicine, Kocaeli University, Kocaeli, Turkey 5 Department of Biochemistry, School of Medicine, Medeniyet University, Istanbul, Turkey Summary Kratak sadr`aj Background: The aim of the study was to investigate the association of paraoxonase 1 (PON1) polymorphism, PON1/ arylesterase (ARE) activity and oxidative stress index (OSI) in breast cancer (BC) patients with type 2 diabetes (DM). Methods: Our study group consisted of 30 healthy women (HV group) and 66 female BC patients. The BC patients were divided into two groups: those with (n=37) and without DM (n=29) (BDM and NBDM group). Genotyping of PON1 Q192R and L55M polymorphisms were done by polymerase chain reaction (PCR) – restriction fragment length polymorphism (RFLP) method. Serum PON1/ARE enzyme activities, total oxidant status (TOS) and total antioxidant status (TAS) were analysed by spectrophotometric method. The ratio of TOS to TAS was accepted as the oxidative stress index (OSI). Results: PON1 Q192R genotype frequency distribution was significantly different in the BDM group compared to the NBDM group (p=0.021). When alleles distribution was examined, R and L alleles were significantly lower, Q and M alleles were significantly higher in the BDM group than in the NBDM group (p<0.001). TOS and OSI were statistically higher in BC patients than HV group (p<0.001). Conclusions: Our results suggest that PON1 gene Q and M alleles may be the risk factors predisposing formation of BC Uvod: Cilj studije bio je istra`ivanje veze polimorfizma paraoksonaze 1 (PON1), aktivnosti PON1/arilesteraze (ARE) i indeksa oksidativnog stresa (OSI) kod pacijentkinja sa rakom dojke (BC) i dijabetesom tip 2 (DM). Metode: Na{a studijska grupa se sastojala od 30 zdravih `ena (HV grupa) i 66 `ena sa rakom dojke (BC). Pacijentkinje iz BC grupe su podeljene u dve grupe: one sa (n = 37) i bez DM (n = 29) (BDM i NBDM grupa). Genotipizacija PON1 K192R i L55M polimorfizama vr{ena je polimeraznom lan~anom reakcijom (PCR) – metodom polimorfizma du`ine restrikcionih fragmenta (RFLP). Enzimske aktivnosti serumske PON1/ARE, ukupni oksidativni status (TOS) i ukupni antioksidativni status (TAS) analizirani su spektrofotometrijskom metodom. Odnos TOS-a prema TAS-u je prihva}en kao indeks oksidativnog stresa (OSI). Rezultati: Frekvencija distribucije genotipova PON1 K192R bila je zna~ajno druga~ija u BDM grupi u odnosu na NBDM grupu (p = 0,021). Kada je ispitana alelna distribucija, R i L aleli su bili zna~ajno ni`i, K i M aleli su bili zna~ajno vi{i u BDM grupi nego u NBDM grupi (p < 0,001). TOS i OSI su bili statisti~ki vi{i kod pacijentkinja sa rakom dojke odnosu na HV grupu (p < 0,001). Zaklju~ak: Na{i rezultati ukazuju na to da PON1 gen Q i M aleli mogu biti faktori rizika koji predisponiraju razvoj raka Address for correspondence: Dr. Fatma Ceyla Eraldemir M.D. Department of Biochemistry School of Medicine, Kocaeli University 41380 Kocaeli, Turkey Tel: +90 262 303 72 56 e-mail: ceyeraldemirªgmail List of abbreviations: BC, breast cancer; PON1, paraoxonase 1; ARE, arylesterase; TOS, total oxidant status; TAS, total antioxidant status; OSI, oxidative stress index; HV, healthy volunteers; BDM, breast cancer with type 2 diabetes mellitus; NBDM, breast cancer without type 2 diabetes mellitus. J Med Biochem 2019; 38 (3) due to increased oxidant damage seen in DM. However, these statements require further confirmation with screening PON1 polymorphism in a greater number of patients with DM, and also wide range follow-up studies are necessary for the same purpose. Keywords: diabetes mellitus type 2, paraoxonase 1, single 369 dojke zbog pove}anog oksidativnog o{te}enja kod DM. Ipak, ovakva tvrdnja zahteva dalju potvrdu skriningom PON1 polimorfizma kod ve}eg broja pacijentkinja sa DM, a u istu svrhu su potrebne i dodatne studije {irokog spektra. Klju~ne re~i: dijabetes melitus tip 2, paraoksonaza 1, jednonukleotidni polimorfizam, oksidativni stres, rak dojke nucleotide polymorphism, oxidative stress, breast cancer Introduction Breast cancer (BC) has become a worldwide public health problem. In Turkey, BC is the most common tumour between women in all age groups as mentioned in Turkey unified database 2013. In 2010, the American Diabetes Association (ADA) and the American Cancer Society (ACS) had a common opinion that diabetic patients have an increased risk of BC (1). Progression of type 2 diabetes (DM) may have a critical role in the initiation and progression of carcinogenesis (2). The mechanisms underlying the relationship between DM and BC may be based on oxidative stress. Oxidative stress may cause cell transformation process raising the risk of cancer development (3). Paraoxonase 1 (PON1) and arylesterase (ARE) are high-density lipoprotein (HDL)-bound enzyme system of antioxidants which protect LDL and HDL from oxidation. Lipid peroxides and hydrogen peroxide are neutralised by PON1 (4). PON1 activity is decreased in diabetic patients (4, 5). Polymorphism in PON1 gene leads to amino acid differences in position 192 of PON1 enzyme (4). Additionally, PON1 enzyme has got ARE enzyme activity and that part of the enzyme is nonpolymorphic. It has been reported that ARE activity is a marker of the actual protein concentration, independent of changes in PON1 activity (6, 7). Loss of the protective effect of PON1 may play an important role in increasing susceptibility to genomic damage and modulate progression of BC causing releasing of some inflammation factors, oxidants and exogenous carcinogens. In addition, there are studies reporting that the activity of PON1 is decreased in DM patients (6, 8). Our aim was to investigate the relationship between PON1 polymorphism, PON1/ARE activity and oxidative stress in patients with DM and BC. Oxidative stress index was used to determine whether PON1 polymorphism could be used as a preliminary diagnostic marker for BC susceptibility of diabetic patients. Materials and Methods A total of 66 patients with BC who underwent surgery and 30 healthy volunteers (HV group) with no family history of BC were enrolled to participa (...truncated)