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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 ADAMTS4 and ADAMTS5 Knockout Mice Are Protected from Versican but Not Aggrecan or Brevican Proteolysis during Spinal Cord Injury(Hindawi Ltd, 2014) Demircan, Kadir; Topcu, Vehap; Takigawa, Tomoyuki; Akyol, Sumeyya; Yonezawa, Tomoko; Ozturk, Gulfer; Ugurcu, VeliThe chondroitin sulfate proteoglycans (CSPGs) aggrecan, versican, and brevican are large aggregating extracellular matrix molecules that inhibit axonal growth of the mature central nervous system (CNS). ADAMTS proteoglycanases, including ADAMTS4 and ADAMTS5, degrade CSPGs, representing potential targets for ameliorating axonal growth-inhibition by CSPG accumulation after CNS injury. We investigated the proteolysis of CSPGs in mice homozygous for Adamts4 or Adamts5 null alleles after spinal cord injury (SCI). ADAMTS-derived 50-60 kDa aggrecan and 50 kDa brevican fragments were observed in Adamts4-/-, Adamts5-/-, and wt mice but not in the sham-operated group. By contrast Adamts4-/- and Adamts5-/- mice were both protected from versican proteolysis with an ADAMTS-generated 70 kDa versican fragment predominately observed in WT mice. ADAMTS1, ADAMTS9, and ADAMTS15 were detected by Western blot in Adamts4-/- mice' spinal cords after SCI. Immunohistochemistry showed astrocyte accumulation at the injury site. These data indicate that aggrecan and brevican proteolysis is compensated in Adamts4-/- or Adamts5-/- mice by ADAMTS proteoglycanase family members but a threshold of versican proteolysis is sensitive to the loss of a single ADAMTS proteoglycanase during SCI. We show robust ADAMTS activity after SCI and exemplify the requirement for collective proteolysis for effective CSPG clearance during SCI.Öğe Caffeic Acid Phenethyl Ester as a Protective Agent against Nephrotoxicity and/or Oxidative Kidney Damage: A Detailed Systematic Review(Hindawi Ltd, 2014) Akyol, Sumeyya; Ugurcu, Veli; Altuntas, Aynur; Hasgul, Rukiye; Cakmak, Ozlem; Akyol, OmerCaffeic acid phenethyl ester (CAPE), an active component of propolis, has been attracting the attention of different medical and pharmaceutical disciplines in recent years because of its antioxidant, anti-inflammatory, antiproliferative, cytotoxic, antiviral, antifungal, and antineoplastic properties. One of the most studied organs for the effects of CAPE is the kidney, particularly in the capacity of this ester to decrease the nephrotoxicity induced by several drugs and the oxidative injury after ischemia/reperfusion (I/R). In this review, we summarized and critically evaluated the current knowledge regarding the protective effect of CAPE in nephrotoxicity induced by several special medicines such as cisplatin, doxorubicin, cyclosporine, gentamycin, methotrexate, and other causes leading to oxidative renal injury, namely, I/R models and senility.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 Female Leuciscus lepidus May Be a New Alternative for Fish Oil Supplements(Hindawi Ltd, 2015) Cakmak, Ozlem; Altuntas, Aynur; Ugurcu, Veli; Erdemli, Hacj Kemal; Akyol, SumeyyaThe proximate composition of male and female Leuciscus lepidus in Beysehir Lake was investigated. The fatty acid profiles of total lipid, phospholipid, and triacylglycerol in muscle and liver of male and female L. lepidus were evaluated by gas chromatography. Proximate analyses showed that meat of male and female L. lepidus had 15.13 +/- 0.04 and 18.75 +/- 0.11% fat, 20.42 +/- 0.45 and 22.21 +/- 0.56% protein, 65.47 +/- 1.37 and 61.28 +/- 1.03% moisture, and 1.51 +/- 0.05 and 1.50 +/- 0.03% ash, respectively. The percentage of total saturated fatty acids was higher in liver than in muscle, whereas the total polyunsaturated fatty acid (PUFA) content was the lowest in all fatty acid profiles. The phospholipids contained more PUFAs than triacylglycerol. Analysis of variance indicated significant differences (P < 0.05) between male (47.51%) and female (49.98%) muscle PUFAs in total lipid. The proportion of omega 3 (omega 3) to omega 6 (omega 6) fatty acids of total lipid was 3.15 in male and 3.68 in female. The ratio is an important indicator for comparing the value of fish oil. Therefore, it was concluded that L. lepidus was considered to be a high quality product for healthy food choice. Additionally, female L. lepidusmay especially be used to produce fish oil supplements from freshwater fish combined with vegetable oils.Öğe Hydrogen Peroxide-Induced Oxidative Damage in Human Chondrocytes: The Prophylactic Effects of Hypericum Perforatum Linn Extract on Deoxyribonucleic Acid Damage, Apoptosis and Matrix Remodeling by a Disintegrin-Like and Metalloproteinase With Thrombospondin Motifs Proteinases(Turkish League Against Rheumatism, 2014) Akyol, Sumeyya; Yukselten, Yunus; Cakmak, Ozlem; Ugurcu, Veli; Altuntas, Aynur; Gurler, Mukaddes; Akyol, OmerObjectives: This in vitro study aimed to examine the protective roles of Hypericum perforatum Linn (HPL) extract on cell viability, DNA damage, apoptosis and a disintegrin-like and metalloproteinase with thrombospondin motifs (ADAMTS) proteins in chondrocytes induced by hydrogen peroxide (H2O2), as a model of chondrocytes subjected to reactive oxygen species attack in rheumatoid arthritis and osteoarthritis. Materials and methods: Human chondrosarcoma cell line (OUMS-27) was used. Cells were incubated with different concentrations of methanolic extract (100, 400, and 750 mu g/ml) of HPL for 36 hours, and then treated with 0.7 mM H2O2 for two hours. Trypan blue was used for evaluation of cell viability, while DNA damage was evaluated by alkaline Comet assay. Caspase-1, ADAMTS5, ADAMTS9, and glyceraldehyde-3-phosphate dehydrogenase proteins were analyzed by Western blot. Results: In vitro H2O2 treatment decreased OUMS-27 cell viability. Cells pretreated with HPL at concentration of 400 mu g/mL were best protected from H2O2 toxicity. Compared to 100 mu g/ml concentration, pretreatment of cells with 750 or 400 mu g/ml of HPL generated more protection against H2O2-induced DNA damage. Hydrogen peroxide application to the cells led to a slight increase in Caspase-1 expression, which shows no apoptosis. The most prominent increase in Caspase-1 level was shown in cells treated with 400 mu g/ml of HPL extract. There was an increase in ADAMTS9 and a decrease in ADAMTS5 levels upon H2O2 administration. Pretreatment with HPL led to more decrease in ADAMTS5 level, indicating the protection of extracellular matrix attacked by these proteinases in cartilage tissue. Conclusion: It can be concluded that HPL has a potential to reverse the negative effects and processes induced by H2O2 in OUMS-27 cells and it can protect the surrounding cartilage area of chondrocytes from oxidative damage, which is suggested to be one of the main molecular factors accused for progression of rheumatoid arthritis and osteoarthritis.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 The Investigation of ADAMTS16 in Insulin-Induced Human Chondrosarcoma Cells(Mary Ann Liebert, Inc, 2015) Cakmak, Ozlem; Comertoglu, Ismail; Firat, Ridvan; Erdemli, Haci Kemal; Kursunlu, S. Fatih; Akyol, Sumeyya; Ugurcu, VeliObjectives: A disintegrin-like metalloproteinase with thrombospondin motifs (ADAMTS) is a group of proteins that have enzymatic activity secreted by cells to the outside extracellular matrix. Insulin induces proteoglycan biosynthesis in chondrosarcoma chondrocytes. The purpose of the present in vitro study is to assess the time course effects of insulin on ADAMTS16 expression in OUMS-27 (human chondrosarcoma) cell line to examine whether insulin regulates ADAMTS16 expression as well as proteoglycan biosynthesis with multifaceted properties or not. Methods: Chondrosarcoma cells were cultured in Dulbecco's modified Eagle's medium having either 10g/mL insulin or not. While the experiment was going on, the medium containing insulin had been changed every other day. Cells were harvested at 1st, 3rd, 7th, and 11th days; subsequently, RNA and proteins were isolated in every experimental group according to their time interval. RNA expression of ADAMTS was estimated by quantitative real-time polymerase chain reaction (qRT-PCR) by using primers. Immunoreactive protein levels were encountered by the western blot protein detection technique by using proper anti-ADAMTS16 antibodies. Results: ADAMTS16 mRNA expression level of chondrosarcoma cells was found to be insignificantly decreased in chondrosarcoma cells induced by insulin detected by the qRT-PCR instrument. On the other hand, there was a gradual decrease in immune-reactant ADAMTS16 protein amount by the time course in insulin-treated cell groups when compared with control cells. Conclusion: It has been suggested that insulin might possibly regulate ADAMTS16 levels/activities in OUMS-27 chondrosarcoma cells taking a role in extracellular matrix turnover.
