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Öğe A disintegrin-like metalloproteinase with thrombospondin motif 8 expression analysis in OUMS-27 chondrosarcoma cells before and after insulin administration(Natl Inst Science Communication-Niscair, 2016) Erden, Gonul; Akyol, Sumeyya; Comertoglu, Ismail; Altuntas, Aynur; Cakmak, Ozlem; Ugurcu, Veli; Yukselten, YunusA disintegrin-like and metalloproteinase with thrombospondin motif 8 (ADAMTS8) is a secreted protease with anti-angiogenic properties. It inhibits vascular endothelial growth factor (VEGF) induced angiogenesis and suppresses fibroblast growth factor 2 (FGF-2) induced vascularization. Angiogenesis and extracellular matrix degradation are the key events in tumor progression, and ADAMTS8 is also known to be a member of the aggrecanases family. In the present study, we investigated the expression levels of ADAMTS8 in chondrosarcoma cells to elucidate the effect of insulin on the tumor cells in terms of ADAMTS production. The OUMS-27 cells were cultured and separately exposed to 10 mu mol/mL insulin up to 11 days in Dulbecco's modified Eagle's medium. After specific time limits (days 1, 3, 7, and 11), the culture was terminated and RNA was isolated using TRIzol reagent and converted to cDNA. The expression levels of ADAMTS8 were evaluated by qRT-PCR. The ADAMTS8 expression in OUMS-27 cells exhibited about 4-fold decrease following insulin treatment on day 11. Statistically significant differences were noted between the control and day 1 (P = 0.008), day 7 (P = 0.047) and day 11 (P = 0.008) groups. The effect of insulin on chondrosarcoma cells in terms of ADAMTS8 expression has not been reported earlier. The decrease in ADAMTS8 expression could be considered as a novel finding that has the potential to explain some pathophysiological mechanisms of tumor cells. Furthermore, the finding could also shed some light on the relationships between matrix degradation and insulin treatment in vitro.Öğe ADAMTS12 Depletion by Insulin in OUMS-27 Human Chondrosarcoma Cells(Turkish League Against Rheumatism, 2015) Altuntas, Aynur; Akyol, Sumeyya; Adam, Bahattin; Cakmak, Ozlem; Ugurcu, Veli; Erden, Gonul; Yukselten, YunusObjectives: In this study, we aim to investigate the association between articular damage in diabetes and a disintegrin-like and metalloproteinase with thrombospondin motifs 12 (ADAMTS12) at gene expression and protein levels. Materials and methods: OUMS-27 human chondrosarcoma cells were used to investigate how ADAMTS12 levels changed in vitro condition in presence and absence of insulin. The study included three groups of cells treated with 10 mu g/mL of insulin, and a control group. Cells were incubated with insulin in medium for one day, three days, and seven days. The effects of insulin on ADAMTS12 were investigated at both gene expression and protein levels. The relationships between the variables were tested by Mann-Whitney U test. Results: ADAMTS12 expression was significantly lower in the groups treated with insulin medium for one day and seven day periods (p=0.008 and p=0.008, respectively) compared to the control group. No significant difference was detected in the expression level between the groups kept in insulin medium for three days and the control group (p=0.55). In addition, protein amounts of the groups exposed to insulin medium for one, three, and seven day periods were lower. Conclusion: Insulin reduces the amount of ADAMTS12 which causes delayed recovery of cartilage tissue in the OUMS-27 cell lines utilized in our study for their chondrocytic properties. This reduction due to insulin treatment may contribute to recovery of cartilage tissue.Öğe ADAMTS13 expression in human chondrosarcoma cells induced by insulin(2014) Fırat, Rıdvan; AKYOL, Sümeyya; KURŞUNLU, S. Fatih; UĞURCU, Veli; Erden, Gonul; ÇAKMAK, Özlem; DEMİRCAN, KadirAmaç: A Disintegrin-like Metalloproteinase with Thrombospondin Motifs (ADAMTS) proteinleri esas olarak ekstraselüler matrikste (ECM) bulunan bir ptoteinaz enzim grubudur. İnsülinin kondrosarkom kondrositlerinde proteoglikan sentezini stimüle ettiği uzun yıllardır bilinmekteydi. Dolayısıyla bu çalışmanın amacı insülinin, OUMS-27 insan kondrosarkom hücre kültüründeki zaman bağımlı etkilerini değerlendirmek ve insülinin anabolik etkilerinden dolayı ADAMTS13 ekspresyonunu azaltma ihtimali hipotezini test etmektir.Yöntemler: Bu hipotezi test etmek için OUMS-27 hücrelerinin, 10?g/mL insulin içeren Dulbecco's modified Eagle besiyerinde (DMEM) kültürü yapıldı. 11. güne kadar iki günde bir besiyeri ile birlikte insülin takviyesi yapıldı. Hücreler 1, 3, 7 ve 11. günlerde harvest edilerek uygun zamanlarda protein ve RNA izolasyonu yapıldı. ADAMTS13 RNA ekspresyonu uygun primerler kullanılarak qRT-PCR yöntemi ile ölçüldü ve anti-ADAMTS13 antikorları kullanılarak Western blot yöntemi ile protein miktarı ölçüldü.Bulgular: Kontrol grubu ile karşılaştırıldığında insülin uygulanan hücre gruplarında, hem RNA hem de protein miktarı azaldı ancak bu istatistiksel olarak anlamlı değildi.Sonuç: Bu bulgular ışığında insülinin OUMS-27 kondrosarkom hücrelerinde ADAMTS13 regülasyonuna pozitif ya da negative herhangi bir etki yapmadığı sonucu ortaya çıkmıştır.Öğe Caffeic Acid Phenethyl Ester: Its Protective Role Against Certain Major Eye Diseases(Mary Ann Liebert, Inc, 2014) Akyol, Sumeyya; Ugurcu, Veli; Balci, Mehmet; Gurel, Ayse; Erden, Gonul; Cakmak, Ozlem; Akyol, OmerAs an effective compound found mainly in the honeybee product propolis, caffeic acid phenethyl ester (CAPE) has been commonly utilized as a medicine and remedial agent, in a number of countries. Specifically, it might inhibit nuclear factor kappa B at micromolar concentrations and demonstrate antioxidant, antineoplastic, antiproliferative, cytostatic, antiviral, antibacterial, antifungal, and anti-inflammatory features. This review article summarizes the recent progress regarding the favorable effects of CAPE on a number of eye disease models, including cataract and posterior capsule opacification, corneal diseases, retina and optic nerve-related diseases, ischemia/reperfusion injury of retina, inflammation and infection-related diseases. CAPE has been found to exhibit promising efficacy, with minimal adverse effects, in animal and cell culture studies of several eye diseases.Öğe Effect of insulin on the mRNA expression of procollagen N-proteinases in chondrosarcoma OUMS-27 cells(Spandidos Publ Ltd, 2015) Akyol, Sumeyya; Comertoglu, Ismail; Firat, Ridvan; Cakmak, Ozlem; Yukselten, Yunus; Erden, Gonul; Ugurcu, VeliChondrosarcoma is one of the most common bone tumors, and at present, there is no non-invasive treatment option for this cancer. The chondrosarcoma OUMS-27 cell line produces proteoglycan and type II, IX, and XI collagens, which constitutes cartilage tissue. A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) proteases are a group of secreted proteases, which include the procollagen N-proteinases ADAMTS-2, -3 and -14. These procollagen N-proteinases perform a role in the processing of procollagens to collagen and the maturation of type I collagen. The present study aimed to improve the understanding of the causes of metastasis, local invasion and resistance to chemo- and radiotherapy in chondrosarcoma, as well as the effect of insulin on cancer cells. The present study was designed to reveal the effects of insulin on procollagen N-proteinases in chondrosarcoma OUMS-27 cells. The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) alone or in DMEM containing 10 mu g/ml insulin. The medium was changed every other day for 11 days. The cells were harvested on days 1, 3, 7 and 11, and total RNA isolation was performed immediately following harvesting. The expression levels of ADAMTS2, ADAMTS3 and ADAMTS14 mRNA were estimated by reverse transcription-quantitative polymerase chain reaction using appropriate primers. ADAMTS2 mRNA expression was found to be decreased on day 7 (P=0.028) and increased at day 11 compared with the control group (P=0.016). The increase in mRNA concentration at day 11 was significantly different compared to the concentrations on days 3 (P=0.047) and 7 (P=0.008). The expression of ADAMTS3 mRNA decreased immediately subsequent to insulin induction on day 1 compared with the control group (P=0.008). The most evident decrease in mRNA concentration was seen at day 7 subsequent to insulin induction (P=0.008). The present results demonstrated that ADAMTS2 and ADAMTS3 may perform a role in the invasion and metastasis of tumors, and may also possess proteolytic activity that results in the breakdown of the extracellular matrix (ECM). Insulin itself can modulate the biosynthesis of ECM macromolecules that are altered in diabetes through various pathways.Öğe Future opportunities in preventing ototoxicity: Caffeic acid phenethyl ester may be a candidate(Spandidos Publ Ltd, 2015) Akyol, Sumeyya; Isik, Bunyamin; Altuntas, Aynur; Erden, Gonul; Cakmak, Ozlem; Kursunlu, Fatih; Adam, BahattinCaffeic acid phenethyl ester (CAPE) is an important active component of propolis, which is derived from honeybee hives. It has received increasing attention in a variety of medical and pharmaceutical research, due to its anti-oxidant, antiproliferative, anti-inflammatory, antiviral and antifungal activity, in addition to its antineoplastic properties. Besides the use of CAPE as an antioxidant and anti-inflammatory agent in a number of in vivo studies of ear disease, its beneficial effects have been reported in the treatment of cancer, arthritis, allergies, heart disease, diabetes, kidney disease, liver disease and neurological disease. CAPE influences a number of biochemical pathways, as well as several targets involved in ear diseases, in particular, in ototoxicity. The protective effects of CAPE in ototoxicity, which may be induced by a number factors, including lipopolysaccharides, hydrogen peroxide and streptomycin, are evaluated and discussed in the present review.Öğe Interleukin-1 induced nuclear factor-B binds to a disintegrin-like and metalloproteinase with thrombospondin type 1 motif 9 promoter in human chondrosarcoma cells(Spandidos Publ Ltd, 2015) Altuntas, Aynur; Halacli, Sevil Oskay; Cakmak, Ozlem; Erden, Gonul; Akyol, Sumeyya; Ugurcu, Veli; Hirohata, SatoshiNuclear factor-B (NF-B) is involved in the regulation of inflammation-associated genes. NF-B forms dimers which bind with sequences referred to as NF-B sites (9-11 bp). A disintegrin-like and metalloproteinase with thrombospondin type 1 motif 9 (ADAMTS9) is a type of proteoglycanase, which proteolytically cleaves versican and aggrecan. ADAMTS9 is a cytokine-inducible gene that contains binding sites for NF-B within its promoter region. Interleukin-1 (IL-1) affects cartilage metabolism and is involved in the NF-B pathway. It is therefore hypothesized that NF-B binding with ADAMTS9 promoters may activate IL-1, thereby promoting chondrocytic cell growth. In the present study, the OUMS-27 chondrocytic human chondrosarcoma cell line was treated with IL-1 with or without inhibitors of NF-B signaling pathways. Chromatin immunoprecipitation (ChIP) and electromobility shift assays (EMSA) were conducted order to analyze the binding of NF-B with the ADAMTS9 promoter region. NF-B-p65 subunit phosphorylation was promoted in IL-1-treated cells, which were not treated with inhibitors of NF-B signaling pathways. By contrast, NF-B-p65 subunit phosphorylation was inhibited in cells that had been treated with BAY-117085, an NF-B pathway inhibitor. ChIP and EMSA assays demonstrated that, following treatment with IL-1, NF-B-p65 bound to elements located at -1177 and -1335 in the ADAMTS9 promoter region, in contrast to the untreated samples. The results of the present study suggested that NF-B may be involved in IL-1-induced activation of ADAMTS9 in human chondrocytes.Öğe Serum vaspin and adiponectin levels in patients with prolactinoma(Taylor & Francis Ltd, 2016) Ginis, Zeynep; Ucar, Fatma; Erdogan, Serpil; Ozturk, Gulfer; Akyol, Sumeyye; Erden, Gonul; Arslan, Muyesser SaykiBackground: Studies investigating serum vaspin and adiponectin levels in patients with prolactinoma are inconclusive. The aim of this study was to evaluate serum vaspin and adiponectin levels in patients with prolactinoma and healthy controls. Methods: A total of 42 prolactinoma patients (Group 1, 21 patients; Group 2, 21 patients) and 30 healthy controls were enrolled in the study. Group 1 consisted of newly diagnosed patients who were never treated or had not received a dopamine agonist (DA) within 6 months prior to screening. Group 2 consisted of prolactinoma patients who were on DA treatment for at least 6 months at the time of screening. The control group (group 3) consisted of healthy controls. Results. Patients with prolactinoma had higher homeostasis model assessment of insulin resistance and lower quantitative insulin sensitivity check index values in comparison to healthy controls (p<0.001 for both). Serum levels of adiponectin and vaspin were also significantly lower in prolactinoma patients when compared to the control group (p<0.01 and p<0.001, respectively). Following adjustment for confounding factors, the respective odds ratios for prolactinoma in patients in the lower subgroup compared with those in the higher subgroup for adiponectin and vaspin were 2.733 (0.621-12.035; p>0.05) and 5.041 (1.191-21.339; p<0.05). Conclusion: This is the first study to demonstrate the presence of low vaspin levels in patients with prolactinomas. Further studies are needed to help establish the roles of vaspin and adiponectin in prolactinoma patients.Öğe Simultaneous analysis of biochemical markers in vitreous humour and serum: a preliminary study on the effect of storage time at -20°C(Taylor & Francis Ltd, 2016) Gurler, Mukaddes; Ozturk, Gulfer; Kir, M. Ziya; Ginis, Zeynep; Erden, Gonul; Akyol, Sumeyya; Kaya, MehmetVitreous humour (VH) is a quite stable and well-protected biological fluid from post-mortem degradation and contamination, making it a usable specimen in forensic pathology. We aimed to analyse the biochemical markers in VH stored for one month at -20 degrees C and to compare them with those of serum samples of the same deceased people. To the best of our knowledge this is the first time this has been reported in the literature. Serum levels of sodium, potassium and calcium, phosphorous and VH levels of calcium, creatinine, and urea were not statistically different between fresh and stored samples. There was no significant difference in VH urea and creatinine levels whereas serum levels were significantly decreased by the 15th and 30th days. In conclusion, urea and creatinine levels in VH showed adequate stability after one month of storage at -20 degrees C, whereas large variations were seen in the stability of inorganic components such as Na, K, Mg, Cl, and P. Our results indicate the validity of using post-mortem VH samples for urea and creatinine but not for VH stored at -20 degrees C for a limited time frame. The differences in sample handling before being stored at -20 degrees C may introduce bias on some components of VH.
