Browsing by Author "Becer, E"
Now showing 1 - 20 of 34
Results Per Page
Sort Options
Item Protective Effects of Citrus Flavonoid Hesperidin in Enterocytes After Induction with TNF-α and IFN-γ Which Mimic the COVID-19 DiseaseÖzgürbüz, U; Vatansever, S; Becer, E; Ensarioglu, HK; Celik, DABACKGROUND/AIMS: Coronavirus disease (COVID-19) is caused by a virus and exhibits various symptoms such as cough, fever, and chills. Flavonoids have a potential inhibitory effect on coronaviruses. In this study, we determined the effects of hesperidin on enterocyte cells (IEC) after tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma induction which mimics the severe acute respiratory therapy-coronavirus-2 (SARS-CoV-2) infection. MATERIALS AND METHODS: The IEC-6 were treated with 50 ng/mL of TNF-alpha and 100 ng/mL of IFN-y for 48 h to mimic inflammatory shock similar to COVID-19 disease. IEC-6 cells were cultured as control, COVID-19 disease mimic, hesperidin prophylactic, or treated groups. The cytotoxicity effect of hesperidin was analyzed using an MTT assay. Serum levels of TNF-alpha and interleukin (IL)8 were evaluated using ELISA. The distributions of TNF-alpha, IFN-y, IL-1ss, Insulin-like growth factor-I, and caspase-3 were analyzed by indirect immunoperoxidase staining. RESULTS: Both TNF-alpha and IL8 levels were higher in TNF-alpha and IFN-y induction of enterocyte culture medium than in the control. Lesser immunoreactivity of TNF-alpha was detected in the treatment group which hesperidin applicate after TNF-alpha and IFN-y combination. While IL-1 immunoreactivity was similar in both the hesperidin prophylactic and treatment groups, lesser immunoreactivity of TNF-alpha was observed in the hesperidin treatment group. Both IFN-y and vascular endothelial growth factor A immunoreactivities were also decreased in the hesperidin treatment group. CONCLUSION: We found that hesperidin had anti-inflammatory and cell protection effects in IEC after TNF-alpha and IFN-gamma induction which mimics the model of SARS-CoV-2 infection. Therefore, hesperidin could be used to reduce gastrointestinal system symptoms in COVID-19 disease.Item Neuroprotective Effects of Hesperidin and Naringin in SK-N-AS Cell as an In Vitro Model for Alzheimer's DiseaseKusi, M; Becer, E; Vatansever, HS; Yücecan, SObjective: Hesperidin and naringin are flavonoids that are found in citrus fruits. Our aim was to create an in vitro model of Alzheimer's disease (AD) and to evaluate the neuroprotective effects of hesperidin and naringin in SK-N-AS and AD model cells. Methods: A beta(25-35) was used to create an AD model in SK-N-AS cells. The cytotoxicity of hesperidin and naringin was evaluated using MTT. beta-amyloid, tau and alpha-synuclein distributions were analyzed using indirect immunoperoxidase staining to investigate the neuroprotective effects of hesperidin and naringin. Results: The AD model was created by 1 mu M of A beta(25-35) for 48 hours after ThT staining. The intensity of beta-amyloid was reduced through both hesperidin and naringin treatment in AD model cells. Both flavonoids significantly decreased the intensity of alpha-synuclein in SK-N-AS and AD model cells. Conclusions: Hesperidin and naringin can be potentially used as neuroprotective agents. Naringin may be more effective than hesperidin in the accumulation of beta-amyloid and tau proteins.Item Quercetin Change the Exosome Secretion and Total miRNA Concentration in Primary (Colo320) and Metastatic (Colo741) Colon Cancer Cell LinesBecer, E; Özsoy, S; Kabadayi, H; Vatansever, HSObjective: Quercetin, which is considered a potential anti-cancer agent in the prevention of colon cancer, is one of its natural polyphenolic compounds. Extracellular vesicles, such as exosomes, secreted from cells and their components contribute to cellular behavioral characteristics by transporting proteins or miRNAs. In this study, we aimed to determine the cytotoxicity of Dicer, Ago2, eIF2 alpha CD9 and CD63 and their effects on exosomal miRNA secretion and expression in Colo320 and Colo741 colon cancer cell lines applied quercetin. Methods: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay was used to analyze the cytotoxicity of quercetin. MTT analysis is a colorimetric analysis method applied to measure the metabolic activity of cells. The absorbance was measured at 570 nm by a spectrophotometer. Besides that, the indirect immunoperoxidase staining was used for the distribution of Dicer, Ago2, eIF2 alpha, CD9, and CD63 in Colo320 and Colo741. Total miRNA in exosome was determined with miRCURYTM Kit. Results: The immunoreactivities of eIF2 alpha and CD9 significantly differed compared to the Colo741 control group after quercetin application. In addition, exosomal miRNA concentrations were higher in both quercetin applied-Colo320 and Colo741 cells. Conclusion: We concluded that quercetin triggered exosomal secretion in Colo320 and Colo741 cells. However, exosomal component should be evaluated in future investigations to understand the quercetin role in primary and metastatic colon adenocarcinoma cells.Item Antiproliferative, antioxidant and anti-inflammatory effects of Boswellia Sacra on human pancreatic cancer cellsBecer, E; Altundag, EM; Özbilenler, C; Vatansever, HS; Baser, KHCPancreatic cancer is a highly lethal and malignant tumor with high occurrence worldwide. Natural products such as essential oils with a lack of side effects may benefit pancreatic cancer supplementary therapy. Essential oil of BS was prepared by distillation of the gum resin and also has been shown to have anti-cancer and antioxidant activities. The present study aimed to determine the anti-cancer, antioxidant and anti-inflammatory activities in PANC-1 cells. Chemical Functional Moieties of Boswellia sacra essential oil were determined by Fourier transform infrared (FTIR) spectroscopy. The essential oil was simultaneously analyzed by GC/MS and GC-FID. Cell viability was determined by MTT assay. Anti-cancer activity of BS essential oil was investigated by immunocytochemistry using antibodies against STAT-3, IHH, beta-catenin, CD133, and Ki-67. The antioxidant activity of BS essential oil was tested by DPPH method. 41 compounds were identified in the essential oil. The IC50 value of BS essential oil was found as 130 mu g/mL in PANC-1 cells. Ki-67 immunoreactivity significantly decreased in BS essential oil-treated PANC-1 cells. STAT-3 and beta-catenin immunoreactivities significantly increased in BS essential oil-treated PANC-1 cells. Also, BS essential oil showed antioxidant and anti-inflammatory activities. We conclude that BS essential oil may suppress proliferation via the beta-catenin activation pathway in PANC-1 cells.Item An Endemic Plant of Cyprus, Origanum majorana: Is It A New Alternative Natural Product for Malaria Treatment?Güler, E; Özbilgin, A; Becer, E; Hanoglu, A; Sanlidag, TMalaria still remains to be a public health threat and one of the most important infectious diseases to get attention from World Health Organization. No domestic malaria cases have been reported on the island of Cyprus since 1948, as a result of successful elimination process. All of the malaria cases detected in recent years are imported cases. As known, hundreds of medicines are obtained from plants and traditional medicine are used in endemic places of malaria. The cause of malaria - Plasmodium parasites, are developing resistance to antimalarial drugs. Hence, research on plant extracts and essential oils have gained great interest in recent years to obtain new and safe agents/substances. In our study, it was aimed to investigate the in vivo antimalarial activities of essential oils obtained from Origanum dubium, Origanum majorana, Salvia fruticosa and Laurus nobilis plants which grows in Northern Cyprus against Plasmodium berghei - the rodent malaria agent. Plants were collected in appropriate seasons and were dried to obtain and analyze essential oils via Clevenger Apparatus system. L929 mouse fibroblast cell line and MTT [3-(4.5-dimethylthiazole-2-yl) -2.5-diphenyltetrazolium bromide] kit were used to determine the cytotoxic activities of the essential oils obtained. In our study, total of 36 mice (Balb/c) of 6 groups (6 mice in each group) were formed: chloroquine group (CG) (50 mg/kg) as malaria reference group, untreated control group (UTCG), O. dubium (OD) (20 mg/kg), O.majorana (OM) (20 mg/kg), S.frutikosa (SF) (20 mg/kg) and L.nobilis (LN) (20 mg/kg). The essential oils were given to mice infected with P.berghei strain orally on 0, 1, 2 and 3rd days (4 times in total). Blood was taken from the tail end of each mouse 24 hours after the last treatment and blood collection was continued every two days until the mice died. Withdrawn blood taken from the mice were prepared as a thin smear and stained with Giemsa. Then, parasitemia percentages in each smear were calculated. As a result of the cytotoxicity tests, cytotoxic activity was not found at 100 mu g/ml (20 mg/kg) in all oils except OD essential oil. While the mice receiving chloroquine continued their lives with the disappearance of the parasite on the 6th day, the mice in the UTCG died on the 9th day. The parasitemia rate reached 35% in the OM group on the 23rd day, in the OD group on the 21st day and in the other groups (SF and LN) on the 14th day and the mice have died. In our study, the difference between the life span in all groups was found statistically significant (ps <= 0.001). As a result, the essential oils O.majorana (14 days increase according to UTCG) an endemic plant of Cyprus and O.dubium (12 days increase according to UTCG) which had an antimalarial effect, decreased parasitemia and increased the life span of mice more than two times, indicated that they could be a source for the acquisition of new antimalarial molecules.Item The role of probiotic content of kefir in the adipogenic differentiation process of 3T3-L1 cellsZeybek, SG; Becer, E; Sanlidag, T; Yücecan, S; Vatansever, SObjective: Kefir is traditional fermented milk that is reported to have various health benefits such as preventing obesity. The aim of this study was to investigate the effect of kefir on lipid accumulation and cell differentiation in adipocytes. Materials and Methods: Kefir was prepared from milk and kefir powder fermentation. Then, kefir supernatant and pellet fractions were incubated for 24 h and 48 h with mature 3T3-L1 adipocytes. Cytotoxic effect of kefir on the cells was measured by MTT assay. Lipid accumulation was detected by Oil-red O staining. Anti-adipogenic activities of kefir fractions were investigated distribution of ANGPTL-4, leptin and PPAR-gamma in mature 3T3-L1 adipocytes using indirect immunoperoxidase technique. Results: In the MTT assay, 0.1 mg/dl dilutions of kefir supernatant and pellet were found to be effective on adipocytes growth and differentiation after both 24 h and 48 h incubations. Lipid accumulation was reduced by both kefir supernatant and pellet in 3T3-L1 adipocytes for 24 h. As a result of immunohistochemical staining, ANGPTL-4 immunoreactivity was significantly increased in 3T3-L1 cells after treated with kefir fractions. Additionally, leptin immunoreactivity was also decreased significantly in the mature adipocytes after treated with kefir pellet. There were no differences in expression of PPAR-gamma in all groups. Conclusion: Kefir fractions have prevented lipid accumulation by preserving the cell structure on 3T3-Li adipocytes. The increased level of ANGPTL4 expression may be mediated inhibition of lipid accumulation. Moreover, kefir supernatant may play important role in preventing weight gain due to reduced leptin level.Item Basic approach on the protective effects of hesperidin and naringin in Alzheimer's diseaseKusi, M; Becer, E; Vatansever, HSObjectivesAlzheimer's disease (AD) is a neurodegenerative disease characterized by cognitive impairment. This situation imposes a great burden on individuals, both economically and socially. Today, an effective method for treating the disease and protective approach to tau accumulation has not been developed yet. Studies have been conducted on the effects of hesperidin and naringin flavonoids found in citrus fruits on many diseases.MethodsIn this review, the pathophysiology of AD is defined, and the effects of hesperidin and naringin on these factors are summarized.ResultsStudies have shown that both components may potentially affect AD due to their antioxidative and anti-inflammatory properties. Based on these effects of the components, it has been shown that they may have ameliorative effects on A beta, alpha-synuclein aggregation, tau pathology, and cognitive functions in the pathophysiology of AD.DiscussionThere are studies suggesting that hesperidin and naringin may be effective in the prevention/treatment of AD. When these studies are examined, it is seen that more studies should be conducted on the subject.Item Composition and antibacterial, anti-inflammatory, antioxidant, and anticancer activities of Rosmarinus officinalis L. essential oilBecer, E; Altundag, EM; Güran, M; Vatansever, HS; Ustürk, S; Hanoglu, DY; Baser, KHCRosemary is a culinary herb that is widely used in traditional medicine for its nutritional value and pharma-cological properties. The aim of this study was to investigate the composition, in vitro anticancer, antioxi-dant, and antibacterial activities of rosemary essential oil. Rosemary essential oil components were simultaneously analysed by GC/MS and GC/FID techniques. Different concentrations of rosemary essential oil were incubated for 24 and 48 h with HepG2 and EV304 cells. Cell viability was measured by MTT assay. Anti-cancer activities of rosemary essential oil were investigated by immunocytochemistry using antibodies directed against Ki-67, b-catenin, c-myc, Oct-3/4, and IL-8. Anti-inflammatory and antioxidant activity were evaluated with protein denaturation and DPPH assays, respectively. The anti-bacterial effect of rosemary essential oil was analysed by Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae with the broth microdilution and agar zone diffusion methods. Camphor (15.1%), verbenone (14.3%), a-pinene (13.6%), 1,8-cineole (11.8%), and borneol (7.9%) were determined as the major compounds of rosemary essential oil. The activities of the oil were found to be 508.7 mg/ml and 525.7 mg/ml against HepG2 and ECV304 cells, respec-tively. Ki-67, b-catenin, c-myc, Oct-3/4, and IL-8 immunoreactivities were significantly reduced in rosemary essential oil-treated HepG2 cells. Ki-67 and b-catenin immunoreactivities were significantly decreased only in rosemary essential oil-treated ECV304 cells. Also, the essential oil showed antioxidant and free radical scavenging activities. Rosemary essential oil showed effective antibacterial activity on E. coli and K. pneumo-niae. Thus, rosemary essential oil could be a potential candidate as a therapeutic agent in cancer treatment.& COPY; 2023 SAAB. Published by Elsevier B.V. All rights reserved.Item Carvacrol is potential molecule for diabetes treatmentHoca, M; Becer, E; Vatansever, HSDiabetes is an important chronic disease that can lead to various negative consequences and complications. In recent years, several new alternative treatments have been developed to improve diabetes. Carvacrol found in essential oils of numerous plant species and has crucial potential effects on diabetes. The anti-diabetic effects of carvacrol have also been comprehensively studied in diabetic animal and cell models. In addition, carvacrol could improve diabetes through affecting diabetes-related enzymes, insulin resistance, insulin sensitivity, glucose uptake, anti-oxidant, and anti-inflammatory mechanisms. The use of carvacrol alone or in combination with anti-diabetic therapies could show a significant potential effect in the treatment of diabetes. This review contributes an overview of the effect of carvacrol in diabetes and anti-diabetic mechanisms.Item Neuroprotective effects of catechins in an experimental Parkinson's disease model and SK-N-AS cells: evaluation of cell viability, anti-inflammatory and anti-apoptotic effectsÖzduran, G; Becer, E; Vatansever, HS; Yücecan, SObjectives The aim of the study was to establish an in vitro Parkinson's disease (PD) model and to investigate the cell viability, anti-inflammatory, anti-apoptotic and neuroprotective effects of catechin and EGCG in SK-N-AS and in vitro PD model cells. Method SK-N-AS human neuroblastoma cells were used. To develop an in vitro PD model, SK-N-AS cells were exposed to 6-hydroxydopamine. Model control was performed after ELISA analysis of dopamine and alpha-synuclein levels in the culture medium. Catechin and EGCG were administered to SK-N-AS and in vitro PD model cells. Cell viability was measured using MTT assay and trypan blue staining. Anti-inflammatory and anti-apoptotic activities of catechin and EGCG were investigated by indirect immunocytochemistry using anti-TNF-alpha, anti-IL-1 beta and anti-caspase-3. Results After 24 hours of 6-hydroxydopamine administration at 50 mu M, higher alpha lfa-synuclein and lower dopamine levels were found in PD model than SK-N-AS cells. Cell viability was similar between SK-N-AS and in vitro PD model cells. Treatment with both bioactive components increased cell viability of in vitro PD model cells. Caspase-3 immunoreactivity was significantly reduced in SK-N-AS and PD model cells after EGCG administration, while it was decreased only in PD model cells after catechin administration. IL-1 beta staining intensity weakened after catechin administration in PD model cells, after EGCG administration in SK-N-AS cells. TNF-alpha staining intensity was similar in both cells. Conclusion Catechin and EGCG increased cell viability in PD model neuron cells. Both components showed anti-apoptotic and anti-inflammatory effects. Catechin may be more effective in preventing damage to neurons PD.Item Cytotoxic activity and antioxidant effects of Origanum onites essential oil and its two major contents, carvacrol and p-cymene on human colorectal (HCT116) and hepatocelluler carcinoma (HepG2) cell linesBecer, E; Altundag, EM; Baser, KHC; Vatansever, HSOriganum onites L. (O. onites) is one of the cultivated Origanum species in Turkey. The aim of this study was to determine the cytotoxicity and cellular antioxidant effects of O. onites essential oil, carvacrol and p-cymene on HCT116 and HepG2 cells. O. onites essential oil components were analyzed by GC/MS and GC/FID. Cytotoxicity was analyzed via the MTT assay in HCT116 and HepG2 cells. Cellular antioxidant activities of O. onites essential oil, carvacrol and p-cymene were determined with DCFH-DA method. GC/MS analysis identified carvacrol (%78.4) as the main constituent of O. onites essential oil. 400 mu g/ml concentration of O. onites essential oil, carvacrol and p-cymene were found to be active against HCT116 and HepG2 cells. Also, carvacrol showed higher cellular antioxidant activity than O. onites essential oil and p-cymene. We conclude that O. onites essential oil and carvacrol may be a candidate for pharmaceutical products in treatment of cancer.Item The effect of Colchicum pusillum in human colon cancer cells via Wnt/β-catenin pathwayBecer, E; Hanoglu, DY; Kabadayi, H; Hanoglu, A; Vatansever, S; Yavuz, DÖ; Meriçli, F; Meriçli, AHObjective: Colchicum pusillum belongs to the family Colchicaceae that particularly rich in tropolonic alkaloids. The aim of this study was to investigate the cytotoxicity and in vitro anticancer activity of Colchicum pusillum ethanolic extract on Colo-320 primer and Colo-741 metastatic colon adenocarcinoma cell lines. Materials and methods: Colchicum pusillum was collected and extracted with ethanol. Different concentrations of Colchicum pusillum extract were incubated for 24 h and 48 h with Colo-320 and Colo-741 cells. Cell growth and cytotoxicity were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Anticancer and antiproliferative activities of Colchicum pusillum were investigated by immunocytochemistry using antibodies directed against to beta-catenin, Ki-67, LGR-5 Ki-67, DKK1, Frizzled-4, Wnt4, Wnt7a and caspase3 in Colo-741 cells. Results: All concentrations of Colchicum pusillum extract had toxic effect in Colo-320 cells. Because of this, we used Colchicum pusillum extract at 20 mu g/ml for evaluate anticancer activities only in Colo-741 cells. As a result of immunohistochemical staining, beta-catenin, LGR-5 and caspase-3 immunoreactivities were significantly increased while Wnt7a immunostaining intensity was decreased in Colo-741 cells. Conclusion We conclude that Colchicum pusillum extract increased beta-catenin and LGR-5 via Wnt/beta-catenin pathway in colon cancer cells. Interestingly, it decreased other signaling molecule, Wnt7a which is assumed to play protective role during carcinogenesis. Also, it increased significantly caspase-3 immunoreactivity showing that apoptotic pathways were triggered.Item The Role and Mechanisms of Action of Catechins in Neurodegenerative DiseasesÖzduran, G; Becer, E; Vatansever, HSThe prevalence, incidence and mortality rates of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease are gradually increasing. New approaches are being developed to manage the progression and treatment of neurodegenerative diseases. Catechins, polyphenolic compounds, are key compounds that demonstrate therapeutic effects with their properties such as antioxidant, anti-inflammatory, anti-apoptotic properties in the prevention and treatment of neurodegenerative diseases. The therapeutic effects of catechins have been exhaustively studied in human and animal models. Catechins can have anti-inflammatory effects by suppressing inflammatory pathways and cytokines, as well as antioxidant effects such as chelating metal ions and scavenging radicals. They might reduce phosphorylation of tau proteins, aggregation of amyloid-beta and apoptotic proteins release. They can also decrease alpha-synuclein accumulation and increase dopamine levels. With all these effects, they can have an effect on neurodegenerative diseases. This review points to the potential mechanisms of catechins in neurodegenerative diseases, based on their findings in the literature review. Key teaching points Catechins can reduce amyloid-beta plaque aggregation and tau phosphorylation. Catechins can decrease alfa-synuclein levels. Catechins can protect neuronal cells with their anti-apoptotic effect. More comprehensive studies are needed to clarify this issue.Item Senescence-mediated anticancer effects of quercetinGokbilen, SO; Becer, E; Vatansever, HSCellular senescence plays a key role in aging and age-related disease initiation. It is a highly dynamic and multistep process that can be stimulated by various stimuli, including cellular stress, DNA damage, telomere shortening, and oncogene activation. Also, senescence is a potent antitumor mechanism, by preventing the proliferation of cancerous cells. However, some of the senescent cells have apoptosis resistance and can cause recurrence in cancer. A new class of drugs termed senolytics selectively kill and eliminate senescent cells. In recent years, natural compounds such as quercetin have been discovered to be effective as senolytic agents. Quercetin is a phytochemical that has strong antioxidant properties and pro-apoptotic effects and has been investigated for many years. Additionally, it has great potential to be used as a senolytic agent. According to preclinical and early-phase clinical data of senolytic agent research, quercetin administration appears to be effective in preventing or alleviating cancer formation. In this paper, we review the importance of cellular senescence in carcinogenesis and the effects of quercetin on senescence, as well as quercetin's potential effects as a pro-apoptotic agent and suppressor of cancer cell proliferation. (c) 2022 Elsevier Inc. All rights reserved.Item Resveratrol modulates miRNA machinery proteins in different types of colon cancer cellsBecer, E; Madencioglu, S; Kabadayi, H; Vatansever, HSObjectives Resveratrol (RSV) is a stilbenoid compound that shows anticancer activity in many cancer cells. Exosomes might affect carcinogenesis and the development of colorectal cancer by affecting communication between tumor cells in the tumor microenvironment via their cargo content miRNA. The aim of this study is to determine the effect of RSV on the expression of Dicer, Ago2, eIf2 alpha, CD-9, CD-63, and exosomal miRNA levels in COLO320 and COLO741 colon cancer cell lines.Methods The MTT method was used for cell growth and cytotoxicity in both COLO320 and COLO741 cell lines. Dicer, Ago2, eIF2 alpha, CD-9, and CD-63 antibodies were used for the immunocytochemical evaluation. Total miRNA analysis was performed using a miRCURY Exosome Isolation Kit.Results As a result of immunocytochemical staining, increased CD-63 immunoreactivity was observed in RSV-treated COLO320 cells vs. RSV-treated COLO-741 cells. Dicer immunoreactivity increased after the RSV treatment in COLO320 cells. Higher eIF2 alpha immunoreactivity was observed in RSV-treated COLO741 cells compared to both COLO741 control cells and RSV-treated COLO320 cells. Non-significant decreases were observed in miRNA concentration in RSV-treated COLO320 and COLO741 cells compared to control group cells.Conclusions RSV could increase miRNA biogenesis in COLO320 cancer cells and decrease it in COLO741 cancer cells.Item Anti-Tumorigenic Effect of Resveratrol in HepG2 cells Controlled with Cytochrome-c Dependent Cell DeathKiyak, N; Becer, E; Vatansever, HS; Kükner, AResveratrol is a phytochemical that is regarded as a potential anticancer agent in liver cancer prevention. It had also been shown that resveratrol has role in the prevention of cancer and anti-cancer properties. In this study, we aimed to investigate the effects of resveratrol on cell viability, apoptosis, cellular proliferation, JAK/STAT pathway and epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma cell (HepG2)line. Cell growth and cytotoxicity were evaluated with MTT assay with different concentrations (5, 10, 25, 50, 100 mu M) of resveratrol in HepG2 cells. The distribution of FasL, cyt-c, caspase-3, Ki-67, ACTA2, CD133, JAK2, N-cadherin, vimentin and STAT3 in HepG2 cells were analyzed using indirect immunoperoxidase technique. The effective dose and incubation time for inhibition of cell growth in HepG2 cells was determined as 100 mu M for 48 hours. Decreased Ki-67 immunoreactivity following resveratrol application was significant in HepG2 cells. Increased cytc-c, STAT3, vimentin, N-cadherin and CD133 immunoreactivities were significant between resveratrol appli-cated HepG2 cells and control group. According to our results, resveratrol induced mitochondrial-dependent cell death and suppressed proliferation in HepG2 cells. On the other hand, our results showed that resveratrol stimu-lated cellular self-protection responses through activation of EMT and STAT3 protein expression in HepG2 cellsItem Obtaining Stem Cell Spheroids from Foreskin Tissue and the Effect of Corchorus olitorius L. on Spheroid ProliferationBecer, E; Soykut, G; Kabadayi, H; Mammadov, E; Çalis, I; Vatansever, SObjectives: Mesenchymal stem cells are self-renewing stem cells. The human foreskin has potential to be used as a source of stem cells. The aim of the study was to obtain spheroid formation of human foreskin cells (hnFSSCs) isolated from newborn human foreskin tissue. In addition, the apoptotic and proliferative effects of a traditional plant, Corchorus olitorius L. (C. olitorius), on hnFSSC spheroids were investigated. Materials and Methods: After a routine circumcision procedure the cells were isolated and cultured in suitable medium. The plant leaves was extracted with ethanol and their composition was analyzed by liquid chromatography coupled with mass spectrometry (LC-MS/MS). The foreskin stem cells were characterized immunocytochemically by CD45, CD34, and CD90 antibodies. hnFSSC spheroids were formed using the hanging drop technique. Immunofluorescence staining was used on the obtained spheroids to determine the distribution of caspase-3 and Ki-67 after being treated with C. olitorius extract for 48 h. Results: Immunostaining analysis showed that hnFSSCs were positive for CD45 and CD34 and negative for CD90. According to LC-MS/MS C. olitorius was rich in flavanols and hydrocinnamic acid derivatives. Although the spheroids obtained were loose and floating, the cells interacted with each other. Caspase-3 activity was higher in the control group than in the extract-treated group and Ki-67 was higher in the extract-treated group than in the control group, suggesting that the plant might have the capacity to increase stem cell proliferation due to its rich polyphenolic content. Conclusion: The results suggest that hnFSSCs and spheroids might be used in stem cell generation, tissue repair and renewal as human foreskin tissue has potential to be used as a stem cell source. C. olitorius also increased proliferation of hnFSSCs, showing that polyphenols might increase proliferation of stem cells.Item Fatty acid composition of Opuntia ficus - indica seed oil control angiogenic activity in colon carcinoma cell linesBecer, E; Kabadayi, H; Meriçli, AH; Kivançli, B; Vatansever, HS; Meriçli, FOpuntia cus-indica belongs to the family Cactaceae that particularly rich in unsaturated fatty acids. The aim of this study was to investigate the composition and in vitro anti-angiogenic activity of spiny and thornless Opuntia ficus-indica seed (CPS) oils. Spiny and thornless CPS oils were obtained by supercritical CO2 extraction and analyzed by GC-MS. Cell growth and cytotoxicity were measured with MTT assay with different concentrations of spiny and thornless CPS oils in Colo-320 and Colo-741 cell lines. Anti-angiogenic activity of CPS oils were investigated by immunocytochemistry using antibodies directed against to Flt-1, Flk-1, eNOS, iNOS, IL-6, PGE(2), TNF-alpha and VEGF. Oleic acid (17.6%; 15.6%), linoleic acid (55.9%; 60.1%), palmitic acid (12.4%; 12.34), elaidic acid (4%; 4.1%) were determined as the major compounds of spiny and thornless CPS oils, respectively. In the MTT assay, 1:16 dilution spiny CPS oil was found to be active against Colo-320 and Colo-741 cells for 48h incubation. Also, 1:8 and 1:16 dilutions of thornless CPS oil were more effective at inhibiting Colo-320 and Colo-741 for 48 h, respectively.We conclude that spiny CPS oil decreased signaling molecules which play in PGE2 mediated and VEGF-dependent angiogenesis. Interestingly, thornless CPS oil increased angiogenesis thought signaling molecules in colon cancer cells.Item The Effect of Resveratrol and Quercetin on Epithelial-Mesenchymal Transition in Pancreatic Cancer Stem CellHoca, M; Becer, E; Kabadayi, H; Yucecan, S; Vatansever, HSResveratrol and quercetin are phytochemicals that are found in a variety of plants. The aim of this study was to investigate the effect of resveratrol and quercetin on epithelial-mesenchymal transition (EMT) of CD133+ and CD133- pancreatic cancer cells. Cancer stem cells (CD133+ cells) were obtained from the PANC-1 cells by the MiniMACS system. CD133+ and CD133- PANC-1 cells were treated with different concentrations (5, 10, 25, 50, and 100 mu M) of resveratrol and quercetin. Cell growth and cytotoxicity were evaluated by MTT assay. Anticancer and anti-metastatic properties of resveratrol and quercetin were determined by immunocytochemistry using antibodies (ACTA-2, IL-1 beta, N-cadherin, TNF-alpha, and vimentin). The immunostaining intensity of CD133+ cells was stronger than CD133- cells. ACTA-2, IL-1 beta, and N-cadherin immunoreactivities were significantly decreased, whereas TNF-alpha and vimentin immunoreactivities significantly increased in quercetin-treated CD133+ cells. Moreover, N-cadherin and TNF-alpha immunoreactivities significantly decreased in resveratrol-treated CD133+ cells. The reduction in N-cadherin and ACTA-2 immunoreactivities was higher than the increase in vimentin immunoreactivity, quercetin could prevent EMT to a greater extent than resveratrol in pancreatic cancer stem cells because of the reduced expression of N-cadherin. Quercetin could be more effective in inhibiting metastasis compared to resveratrol.Item Apoptotic effects of Corchorus olitorius L. leaf extracts in colon adenocarcinoma cell linesSoykut, G; Becer, E; Calis, I; Yucecan, S; Vatansever, SCorchorus olitorius L. is a highly consumed plant in Cyprus and other Eastern Mediterranean countries and known as a medicinal food in many cultures. The aim of the study is to determine in vitro anticancer and apoptotic induction effects of dichloromethane (DCM) and aqueous Con-horus olitorius L. leaf extracts in primary (Colo-320) and metastatic (Colo-741) colon adenocarcinoma cell lines.. Cell growth and cytotoxicity were measured with MTT assay with five different concentrations of extracts in Colo-320 and Colo-741 cell lines. Apoptotic activities of Corchorus olitorius L. were investigated by immunocytochemistry using antibodies directed against to caspase-3, cytochrome-c and FasLigand (FasL). TUNEL assay was used to detect DNA fragmentation in both cell lines. Both DCM and aqueous phase of extracts at 50 mu g/ml concentration were more effective at inhibiting Colo-320 and Colo-741 cell growth when compared with other dilutions. The number of TUNEL positive cells was significantly higher in Colo-741 cells when compared with both control group and Colo-320 cell line. DCM phase extract significantly increased caspase-3 immunoreactivity while aqueous phase extract significantly increased cytochrome-c and FasLigand (FasL) immunoreactivities in Colo-320 cell lines. Both extracts were able to significantly increase caspase-3, cytochrome-c and FasL immunoreactivity in Colo-741 cells showing that both apoptotic pathways were triggered. Overall, Corchorus olitorius L. extracts induced apoptosis in both cancer cell lines while being more effective in metastatic colon adenocarcinoma cell lines suggesting that the extracts might have potential anticancer effects and possibility to be used as precursor to phytomedicinal colon cancer treatment as oppose to chemotherapy.