ANTIPROLIFERATIVE AND APOPTOTIC EFFECTS OF THE ESSENTIAL OIL OF ORIGANUM ONITES AND CARVACROL ON HEP-G2 CELLS

The essential oil Origanum onites L. and its phenolic constituent carvacrol were examined for their cytotoxic and apoptotic effects in a human hepatocellular carcinoma cells Hep-G2. WST-1 and neutral red uptake assays were performed to determine the inhibitory effects of the oil and carvacrol on the growth of the cells. Possible induction of apoptosis by Origanum oil and carvacrol was further investigated by acridine orange/ethidium bromide (AO/EB) staining. Results showed that the Ori- ganum oil and carvacrol was significantly cytotoxic and induced apoptosis in Hep-G2 cells. IC₅₀ value of essential oil and carvacrol was found about 0,009% (v/v) and 500 μM, respectively. After incuba- tion of the cells with Origanum oil and carvacrol, characteristics of apoptotic morphology such as chromatin condensation, shrinkage of the cells and cytoplasmic blebbing was observed. In conclusion, both essential oil and its major constituent carvacrol significantly exhibited cytotoxic and apoptotic activities in hepatocellular carcinoma cells, indicating its potential for use as an anticancer agent

Yaşam Bilimleri ve Biyoteknoloji

Origanum onites L. uçucu yağı ve onun fenolik bileşeni olan karvakrolün insan kanserli karaciğer hücreleri Hep-G2 üzerindeki sitotoksik ve apoptotik etkileri çalışılmıştır. Hücrelerin çoğalması üzerine origanum uçucu yağının ve karvakrolün engelleyici etkilerini belirlemek için WST-1 ve nötral kırmızısı alım yöntemleri uygulanmıştır. Origanum yağı ve karvakrolün olası apoptotik etkisi akridinoranj/etidyumbromür (AO/EB) boyama yöntemiyle araştırılmıştır. Sonuçlar değerlendirildiğinde, Origanum yağı ve karvakrolün Hep-G2 hücrelerinde anlamlı derecede sitotoksik etkiye sahip olduğu ve bu hücrelerde apoptozu uyardığı gözlenmiştir. Uçucu yağın IC50 değeri % 0.009, karvakrolün 500 mM olarak belirlenmiştir. Origanum uçucu yağı ve karvakrol ile inkübasyonun ardından hücrelerin kromatin yoğunlaşması, hücre büzülmesi, sitoplazmik keselerin oluşumu gibi karakteristik apoptoz morfolojisi gösterdiği belirlenmiştir. Sonuç olarak, hem Origanum uçucu yağı hem de onun temel bileşeni olan karvakrol, insan kanserli karaciğer hücrelerinde anlamlı bir sitotoksik etki ve apoptotik aktivite sergilemiştir ve bu özelliklerinden dolayı, her ikisi de antikanser ajan olarak kullanılabilme potansiyeline sahipti

Yaşam Bilimleri ve Biyoteknoloji

Origanum onites L. uçucu yağı ve onun fenolik bileşeni olan karvakrolün insan kanserli karaciğer hücreleri Hep-G2 üzerindeki sitotoksik ve apoptotik etkileri çalışılmıştır. Hücrelerin çoğalması üzerine origanum uçucu yağının ve karvakrolün engelleyici etkilerini belirlemek için WST-1 ve nötral kırmızısı alım yöntemleri uygulanmıştır. Origanum yağı ve karvakrolün olası apoptotik etkisi akridinoranj/etidyumbromür (AO/EB) boyama yöntemiyle araştırılmıştır. Sonuçlar değerlendirildiğinde, Origanum yağı ve karvakrolün Hep-G2 hücrelerinde anlamlı derecede sitotoksik etkiye sahip olduğu ve bu hücrelerde apoptozu uyardığı gözlenmiştir. Uçucu yağın IC50 değeri % 0.009, karvakrolün 500 mM olarak belirlenmiştir. Origanum uçucu yağı ve karvakrol ile inkübasyonun ardından hücrelerin kromatin yoğunlaşması, hücre büzülmesi, sitoplazmik keselerin oluşumu gibi karakteristik apoptoz morfolojisi gösterdiği belirlenmiştir. Sonuç olarak, hem Origanum uçucu yağı hem de onun temel bileşeni olan karvakrol, insan kanserli karaciğer hücrelerinde anlamlı bir sitotoksik etki ve apoptotik aktivite sergilemiştir ve bu özelliklerinden dolayı, her ikisi de antikanser ajan olarak kullanılabilme potansiyeline sahipti

Identification of biomarkers associated with partial epithelial to mesenchymal transition in the secretome of slug over-expressing hepatocellular carcinoma cells

Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths worldwide. Complete epithelial to mesenchymal transition (EMT) has long been considered as a crucial step for metastasis initiation. It has, however, become apparent that many carcinoma cells can metastasize without complete loss of epithelial traits or with incomplete gain of mesenchymal traits, i.e., partial EMT. Here, we aimed to determine the similarities and differences between complete and partial EMT through over-expression of the EMT-associated transcription factor Slug in different HCC-derived cell lines

Effects of fibronectin and type IV collagen on osteosarcoma cell apoptosis

789-796The aims of this study are the investigation of the effects of fibronectin and type IV collagen extracellular matrix proteins and the role of caspase-3 and -9 on cis-platin induced U2-OS apoptosis were studied. First the cytotoxic effects of cis-platin on cell system were investigated by colorimetric method and than morphological and ELISA analysis were used for determination of cell apoptosis when induced with cis-platin. In addition, after adhering the cells to fibronection or type IV collagen proteins, the apoptotic rate and the effects of caspase-3 and -9 were also investigated by ELISA in presence of specific inhibitors. U2-OS cells showed 20% cytotoxicity after treatment with 2.4 µM of cis-platin for 48 h. Morphological and the numerical data showed that cis-platin was able to induced apoptosis on cells as a dose-dependent manner. Caspase-3 and -9 inhibitors inhibited cis-platin-induced apoptosis in U2-OS cells, respectively. The binding of cells to 10 µg/mL of fibronectin but not type IV collagen enhanced the apoptosis about 2.5 fold that effects inhibited with caspase-3 inhibitor. The caspase-3 and -9 are involved in the apoptotic signals induced by cis-platin in U2-OS. The binding to fibronectin, but not type IV collagen enhanced the apoptotic response of U2-OS and fibronectin-dependent apoptosis was activated by caspase-3. These finding might be useful for patients to fight against osteosarcoma

ZnO microparticle-loaded chitosan/poly(vinyl alcohol)/acacia gum nanosphere-based nanocomposite thin film wound dressings for accelerated wound healing

WOS:000484279500001The present study deals with the development of novel ZnO microparticle-loaded chitosan/poly(vinyl alcohol)/acacia gum nanosphere-based nanocomposite thin films through electrospraying and evaluation of their potential use in wound healing applications for skin. ZnO microparticles were synthesized and used as bioactive agents. Morphology, size distribution, structure, and dispersion of the synthesized ZnO microparticles were analyzed by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and transmission electron microscopy (TEM). ZnO microparticles were incorporated into the ternary nanocomposite films by electrospraying technique. Thermogravimetric analyses reveal that incorporation of ZnO microparticles into the nanocomposite structure improves the thermal stability. Mechanical analyses show that tensile strength reaches to the maximum value of 12.75 MPa with 0.6 wt % ZnO content. SEM and TEM micrographs demonstrate that the nanocomposite films consist of nanospheres with nanocapsular structures whose sizes are mostly between 250 and 550 nm. Viability tests established prevailing cellular performance of the fibroblasts on 0.6 wt % ZnO microparticle-loaded nanocomposite films with a viability percentage of 160% compared to the control group. (c) 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019, 136, 48445.Anadolu University Scientific Research Projects CommitteeAnadolu University [1705F237]This study was funded by the Anadolu University Scientific Research Projects Committee; Project No: 1705F237. The authors thank Mr. Metin Cam, electrical engineer in Department of Electrical and Electronics Engineering, Anadolu University, for his special efforts in constructing the electrospraying system

Stimulation of dendritic cells with vaccine and vaccine–antibody complex and effect on immune response

İstanbul Bilim Üniversitesi, Sağlık Hizmetleri Meslek Yüksekokulu.Dendritic cell (DC) vaccines are a promising and potent therapeutic tool for chronic diseases, autoimmune diseases, and cancer because of the unique ability of DCs to stimulate T cells. The challenge of DC vaccines is to find an effective form for antigen presentation. Although pure antigens, antigen complexes, plasmids, and mRNA have been used in different studies, no proper application to overcome this problem has been found yet. In this study, we investigated the eligibility of a commercial hepatitis B virus (HBV) vaccine or a vaccine–monoclonal antibody complex for antigen loading of DCs for a therapeutic purpose. DCs were derived from the bone marrow of transgenic hepatitis B (HBV-tg) mice using a granulocyte macrophage-colony stimulating factor and interleukin-4, and then loaded with a commercial HBV vaccine (containing hepatitis B virus surface antigens and aluminum hydroxide adjuvant) or a vaccine–antibody complex. HBV-tg mice were immunized with the vaccine and vaccine–antibody loaded DCs. Optimum HBV vaccine concentration and loading time were determined by 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium (WST- 1) methods. Therapeutic effects of vaccine–antibody loaded DCs were determined by the evaluation of antibody response and hepatitis B surface expression levels in HBV-tg mice. Our results showe

Cytotoxicity of the methanol extracts of Elephantopus mollis, Kalanchoe crenata and 4 other Cameroonian medicinal plants towards human carcinoma cells

Abstract Background Cancer still constitutes one of the major health concerns globally, causing serious threats on patients, their families, and the healthcare system. Methods In this study, the cytotoxicity of the methanol extract of Elephantopus mollis whole plant (EMW), Enantia chlorantha bark (ECB), Kalanchoe crenata leaves (KCL), Lophira alata bark (LAB), Millettia macrophylla leaves (MML) and Phragmanthera capitata leaves (PCL) towards five human solid cancer cell lines and normal CRL2120 fibroblasts, was evaluated. Extracts were subjected to qualitative chemical screening of their secondary metabolite contents using standard methods. The cytotoxicity of samples was evaluated using neutral red uptake (NR) assay meanwhile caspase activation was detected by caspase-Glo assay. Flow cytometry was used to analyze the cell cycle distribution and the mitochondrial membrane potential (MMP) whilst spectrophotometry was used to measure the levels of reactive oxygen species (ROS). Results Phytochemical analysis revealed the presence of polyphenols, triterpenes and sterols in all extracts. The IC50 values of the best samples ranged from 3.29 μg/mL (towards DLD-1 colorectal adenocarcinoma cells) to 24.38 μg/mL (against small lung cancer A549 cells) for EMW, from 2.33 μg/mL (mesothelioma SPC212 cells) to 28.96 μg/mL (HepG2 hepatocarcinoma) for KCL, and from 0.04 μg/mL (towards SPC212 cells) to 0.55 μg/mL (towards A549 cells) for doxorubicin. EMW induced apoptosis in MCF-7 cells mediated by MMP loss and increased ROS production whilst KCL induced apoptosis via ROS production. Conclusion This study provides evidences of the cytotoxicity of the tested plant extract and highlights the good activity of Elephantopus mollis and Kalanchoe crenata. They deserve more exploration to develop novel cytotoxic drugs

Biological assays on the effects of Acra3 peptide from Turkish scorpion Androctonus crassicauda venom on a mouse brain tumor cell line (BC3H1)

İstanbul Bilim Üniversitesi, Sağlık Hizmetleri Meslek Yüksekokulu.Constitutes of the venom scorpion are a rich source of low molecular mass peptides which are toxic to various organisms, including man. Androctonus crassicauda is one of the scorpions from the Southeastern Anatolia of Turkey with public health importance. This work is focused on the investigation of biological effects of Acra3 peptide from Androctonus crassicauda. For this purpose, Acra3 isolated from crude venoms was tested for its cytotoxicity on BC3H1 mouse brain tumor cells using tetrazolium salt cleavage and lactate dehydrogenase activity assays. To determine whether the cytotoxic effects of Acra3 was related to the induction of apoptosis, the morphology of the cells and the nuclear fragmentation was examined by using Acridin Orange staining and DNA fragmentation assay, respectively. Caspase 3 and caspase 9 activities were measured spectrophotometrically and flow cytometric assay was performed using Annexin-V FITC and Propidium Iodide staining. Furthermore toxic peptide Acra3 was used as an antigen for immunological studies. Results showed that Acra3 exerted very strong cytotoxic effect on BC3H1 cells with an IC50 value of 5 μg/ml. Exposure of the cells to 0.1 and 0.5 μg/ml was resulted in very strong appearance of the apoptotic morphology in a dose dependent manner. On the other side, not any DNA fragmentation was observed after treatment of the cells. Caspase 3 and 9 activities were slightly decreased with Acra3. Results from flow cytometry and lactate dehydrogenase activity assays indicate that Acra3 exerts its effects by inducing a stronger necrosis than apoptosis in BC3H1 cells. To evaluate its immunogenicity, monoclonal antibody (MAb) specific for Acra3 antigen (5B9) was developed by hybridoma technology using spleen and lymph nodes of mice and immunoglobulin type of antibody was found to be IgM. We suggest that Acra3 may exert its effects by inducing both necrotic and apoptotic pathway in some way on mouse brain tumor cells. These findings will be useful for understanding the mechanism of cell death caused by venom in vitro. Anti-Acra3 monoclonal antibody can be further used as a bioactive tools for exploring the structure/function relationship and the pharmacological mechanism of scorpion peptide neurotoxins