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Öğe Analysis of MMP-7 and TIMP-2 gene polymorphisms in coronary artery disease and myocardial infarction: A Turkish case-control study(Elsevier (Singapore) Pte Ltd, 2017) Alp, Ebru; Yılmaz, Akın; Tulmaç, Murat; Uğraş Dikmen, Asiye; Çengel, Atiye; Yalçın, Rıdvan; Menevşe, Emine SevdaMatrix metalloproteinase (MMP) and tissue inhibitors of metalloproteinase (TIMP) have a significant role in tissue remodeling related to cardiac function. In earlier studies, MMP-7 A-181G (rs11568818), C-153T (rs11568819), C-115T (rs17886546), and TIMP-2 G-418C (rs8179090) polymorphisms have been studied in various diseases. However, association between coronary artery disease (CAD) and these polymorphisms has been poorly studied. The goal of this study is to investigate the association of CAD and myocardial infarction (MI) with MMP-7 or TIMP-2 polymorphisms. This study included 122 CAD patients and 132 control individuals. DNA was extracted from whole blood. Polymerase chain reaction-restriction fragment length polymorphism and automated direct sequencing method were used for genotyping of these polymorphisms. No significant differences were found between MMP-7 A-181G, C-115T, and TIMP-2 G-418C polymorphism and CAD or MI in a Turkish population. Despite the fact that the genotypes of MMP-7 C-153T polymorphism had no significant differences among MI and control groups, allele frequencies of C-153T polymorphism were significantly different between the two groups. Our study is the first report to clarify the appreciable relationship between MMP-7 C-153T polymorphism and MI development in CAD patients. However, these findings also need to be confirmed in other populations so we can improve our knowledge about the genetic factors affecting the development of CAD. © 2017Öğe Carboxylesterase 1, alpha 2a adrenergic receptor and noradrenalin transporter gene polymorphisms and their clinical effects in attention deficit hyperactivity disorder in Turkish children(ScienceDirect, 2018-06) Çetin, Fatih Hilmi; Işık, Yasemen; Alp, Ebru; Yılmaz, Akın; Taş Torun, Yasemen; Önen, İlke HacerThe objective of this study was to examine the association between ADHD and G1287A polymorphism in the NET1 gene, C1291G polymorphism in the ADRA2A gene on the adrenergic pathway and Gly143Glu polymorphism in the CES1 gene on the metabolic pathway, and their clinical effects. The study population included 114 children with ADHD and 83 healthy controls. 103 patients are followed for 6?months, their scale points are recorded and side effects are questioned in each interview. Every patient in both control and ADHD group are found to have GG genotype when Gly143Glu polymorphism in the CES1 gene is examined, thus we came to a conclusion that Turkish population is homozygote in the mentioned polymorphism. No significant association between NET1 gene G1287A polymorphism genotypes and ADHD was found. It was found that ADRA2A C1291G polymorphism C allele and CC genotype is a risk factor for ADHD (p?=?0.003, OR: 2.17, CI: 12.8–37.0) and the risk is higher in males (p?=?0.013, OR: 2.43, CI: 12.0–49.5). There was no significant relation between ADRA2A C1291G polymorphism and clinical parameters but it was found that individuals with NET1 G1287A polymorphism AA genotype have less concurrent Oppositional Defiant Disorder diagnosis (18.8% vs. 81.2%, p?=?0.039), their initial CTRS-attention deficit points are higher (17.47?±?3.73 vs.16.15?±?4.58, p?=?0.045). In conclusion, our study showed that the ADRA2A C1291G polymorphism C allele and CC genotype is associated with ADHD. NET1 G1287A polymorphism AA genotype is mainly associated with attention deficit cluster.Öğe Effects of EF-24, RAD001, and paclitaxel on the expression profiles of apoptotic and anti-apoptotic genes(Wolters Kluwer - Medknow, 2017) Alp, Ebru; Yılmaz, Akın; Önen, İlke Hacer; Menevşe, Emine SevdaContext: Cancer cells exert differential responses to chemotherapeutics and inhibitors. To the best of our knowledge, a few or no research has been performed until now to determine the effect of EF-24 and RAD001 on MDA-MB-231 breast cancer cells with regard to mRNA expression of apoptotic and anti-apoptotic genes. Aims: In this study, we aimed to investigate the mRNA expression levels of apoptotic (caspase 2 [CASP2], CASP8, and CASP9) and anti-apoptotic (B-cell lymphoma 2 [BCL2] and BCL2-like protein 1 [BCL2L1]) genes after exposure to paclitaxel, EF-24, and RAD001 in MDA-MB-231 cells. Materials and Methods: After treatment, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to measure cell viability. mRNA expressions were analyzed using quantitative real-time polymerase chain reaction. Results: Decrease in cell viability ratios was seen in a dose-dependent manner for all chemicals. MDA-MB-231 cells responded slightly different to paclitaxel, EF-24, and RAD001 at the transcriptional level of apoptotic and anti-apoptotic genes. Conclusions: Our results showed that response of these cells to paclitaxel, EF-24, and RAD001 was found different at the transcriptional level of apoptotic and antiapoptotic genes. Therefore, understanding transcriptional changes after these drug exposure may give us a change to figure out more realistic results of the apoptotic pathway inhibition. © 2017 Indian Journal of Ophthalmology | Published by Wolters Kluwer - Medknow.Öğe HDAC inhibitors, MS-275 and salermide, potentiates the anticancer effect of EF24 in human pancreatic cancer cells(Leibniz Research Centre for Working Environment and Human Factors, 2016) Yar Sağlam, Atiye Seda; Yılmaz, Akın; Önen, İlke Hacer; Alp, Ebru; Kayhan, Handan; Ekmekci, AbdullahHistone deacetylases (HDACs) play a major role in the regulation of chromatin structure and gene expression by changing acetylation status of histone and non-histone proteins. MS-275 (entinostat, MS) is a well-known benzamide- based HDACI and Salermide (SAL), a reverse amide compound HDACI, have antiproliferative effects on several human cancer cells. In this study, we aimed to investigate the effects of HDACIs (MS and SAL) alone and/or combined use with EF24 (EF), a novel synthetic curcumin analog, on human pancreatic cancer cell line (BxPC-3). In vitro, BxPC-3 cells were exposed to varying concentrations of MS, SAL with or without EF, and their effects on cell viability, acetylated Histone H3 and H4 levels, cytotoxicity, and cleaved caspase 3 levels, and cell cycle distribution were measured. The viability of BxPC-3 cells decreased significantly after treatment with EF, MS and SAL treatments. MS and SAL treatment increased the acetylation of histone H3 and H4 in a dose dependent manner. MS and SAL alone or combined with EF were increased the number of cells in G1 phase. In addition, treatment with agents significantly decreased the ratio of cell in G2/M phase. There were significant dose-dependent increases at cleaved Caspase 3 levels after MS treatment but not after SAL treatment. Our results showed that HDAC inhibitors (MS and SAL), when combined with EF, may effectively reduce pancreatic cancer cell (BxPC-3) progression and stop the cell cycle at G1 phase. Further molecular analyses are needed to understand the fundamental molecular consequences of HDAC inhibition in pancreas cancer cells. © 2016, Leibniz Research Centre for Working Environment and Human Factors. All rights reserved.Öğe HeLa hücrelerinde Sisplatinin MTOR, AKT, CCND1 ve STAT3 mRNA ifadesi üzerine etkileri(Gazi Üniversitesi, 2020) Alp, Ebru; Yılmaz, Akın; Önen, Hacer İlke; Konaç, Ece; Ekmekçi, Abdullah; Menevşe, Emine Sevda; Menevşe, AdnanGiriş: Serviks kanseri, kadınlarda gözlenen malign tümörler arasında en yaygın görülen ve kadınlarda kanser sonucu gözlenen ölümler arasında ikinci sırada bulunan önemli bir hastalıktır. Platin temelli bir bileşik olan sisplatin (CDDP) bu kanserin tedavisinde sıklıkla kullanılan kemoterapötik ajanlardan birisidir ve sarkomlar ile solid tümörlerin tedavisinde etkili bir şekilde kullanılmaktadır. CDDP DNA’ya çapraz bağlanıp replikasyonu engelleyerek hasar oluşumuna neden olur. Bunun sonucunda da apoptotik yolaklar aktive olur ve hücre ölümü gerçekleşir. Çalışmamızda CDDP’nin, mTOR, AKT, Siklin D1 (CCND1) ve STAT3 genlerinin mRNA ifadesindeki etkisini serviks kanseri hücre hattı HeLa üzerinde araştırmayı amaçladık. Yöntem: Bu çalışmada HeLa hücrelerine 24 ve 48 saat süre ile değişen konsantrasyonlarda sisplatin uygulandı. Hücre canlılığı XTT yöntemi ile belirlendi. Ayrıca, seçilen CDDP dozlarının uygulanmasından sonra, mTOR, AKT, CCND1 ve STAT-3 genlerinin mRNA ifadesi qPCR yöntemi ile araştırıldı. Bulgular: CDDP’nin IC50 değerinin 24 saat için yaklaşık 60 uM, 48 saat için ise yaklaşık 8 uM olduğunu belirledik. Ayrıca, analizi yapılan genlerin ifadelerinin sadece 24 saat uygulamadan sonra azaldığını bulduk. Diğer taraftan, 48 saat CDDP uygulamasının mRNA ifadesi açısından istatistiksel açıdan anlamlı bir farkılılığa neden olmadığını gösterdik. Sonuç: Sonuç olarak, CDDP uygulamasından sonra, uygulama süresi açısından değişen, farklı mRNA ifade örüntülerinin olduğunu bulduk. Konvensiyonel kemoterapötik olan CDDP’nin hücre sağkalım yolakları üzerindeki etkisinin ortaya çıkarılması açısından çalışmamız literatüre katkı sağlamaktadır.Öğe Reduced BCL2 and CCND1 mRNA expression in human cervical cancer HeLa cells treated with a combination of everolimus and paclitaxel(Termedia Publishing House Ltd., 2016) Yılmaz, Akın; Alp, Ebru; Önen, İlke Hacer; Menevşe, Emine SevdaAim of the study: Cervical cancer is the second most common malignancy in women worldwide. Everolimus displays direct effects on growth and proliferation of cancer cells via inhibition of mammalian target of rapamycin (mTOR) protein, which is known to be associated with drug resistance. In this study, we aimed to investigate the effects of everolimus, gemcitabine, and paclitaxel in terms of cell viability and mRNA expression levels of GRP78, CCND1, CASP2, and BCL2 genes. Material and methods: HeLa cells were treated with different doses of everolimus, gemcitabine, and paclitaxel. Cell viability was assessed using MTT assay, and obtained dose response curves were used for the calculations of inhibitory concentration (IC) values. At the end of the treatment times with selected doses, RNA isolation and cDNA synthesis were performed. Finally, GRP78, CCND1, CASP2, and BCL2 genes mRNA expression levels were analysed using quantitative PCR. Results: The IC50 value of everolimus was 0.9 M for 24-hour treatment. Moreover, the IC50 value of gemcitabine and paclitaxel was found to be around 18.1 M and 7.08 M, respectively. Everolimus, gemcitabine, and paclitaxel treatments alone did not change the GRP78, CCND1, BCL2 and CASP2 mRNA expression levels significantly. However, combined treatment of everolimus and paclitaxel significantly reduced BCL2 and CCND1 mRNA expression (p < 0.05). In contrast, this combination did not change GRP78 and CASP2 mRNA expression levels (p > 0.05). Conclusions: Down-regulation of CCND1 and BCL2 expression may be an important mechanism by which everolimus increases the therapeutic window of paclitaxel in cervical cancers.
