Kaspaz yolağı ile yönlendirilen insan osteojenik sarcoma hücre çoğalmasına Propolisin baskılayıcı etkisi

Doğal bir ürün olarak propolis belli ağaç ve bitkilerin kabuki ve tomurcuklarından arılar aracılığı ile elde edilen bir reçine materyalidir. Propolis antimutajenik, antioksidan, antibakteriyel, antiviral etkileride içeren biyolojik aktiviteleri olan çeşitli kimyasal bileşikleri içerir. Bu nedenle bu çalışmada insan osteojenik sarcoma hücre dizini SAOS-2 kültüründe kaspaz yolağı kullanılarak propolis ekstraktlarının (PE) antiapoptotik etkisi araştırılması amaçlandı. Ankara’da Hacettepe Üniversitesi Beytepe Kampüs alanında ekolojik çevrede üretilen ekstraktlar kullanıldı. 0.5, 0.25, 0.125 ve 0.063 mg/ml konsantrasyonlarda 7 farklı PE SAOS-2 hücre dizinine eklenerek 2 gün boyunca inkübe edildi. Hücre çoğalması ve toksisitesi için MTT testi ile apoptotic hücre ölümünün belirlenmesi için TUNEL metodu ve kaspaz dağılımı için kaspaz 6, 8 ve 9 immunohistokimyası analizi kullanıldı. Hücreler MTT analizi sonrasında en büyük etki PE 7 için 0.125 mg/ml dilüsyonunda görüldü. TUNEL pozitif hücre sayısı en fazla olan PE 4 ve 5 için 0.125 mg/ml dilüsyonu oldu. Kaspaz 6 immünoreaktivitesi kaspaz 8 ve 9’dan daha güçlü idi. Daha da önemlisi kaspaz 6 boyaması PE 7 için 0.125 mg/ml dilüsyonunda çok daha iyi idi. Sonuç olarak, PE ile indüklenen apoptosis mekanizmasının antikansorejenik etkisinden dolayı kaspaz yolağı ile oluşabilir. PE kullanımı kanser tedavi protokolünde yararlı olabilir.A natural product Propolis, is a resinous material gathered by honeybees from the buds and bark of certain trees and plants. Propolis contains various chemical components of biological activities, including antimutagenic, antioxidant, antibacterial, antiviral and anticarsinogenic. Therefore, the aim of this study is to investigate the antiapoptotic effect of propolis extracts (PE) using caspase pathway in the human osteogenic sarcoma cell line SAOS-2 in culture. The extracts which produced in ecologic environment were taken from the Hacettepe University, Beytepe Campus area-Ankara were used. Seven different PE at 0.5, 0.25, 0.125 and 0.063 mg/ml were added to SAOS-2 cell line for two days incubation. For cell proliferation and cytotoxicty analyses MTT, for apoptotic cell death determination TUNEL method, for distribution of caspase 6, caspase 8 and caspase 9 indirect immunocytochemistry analyses were used. After MTT analyses, the most effect was observed PE 7 at the 0.125 mg/ml dilution. The number of TUNEL positive cells was more detectable at PE 4 and 5 at the 0.063 mg/ml, and PE 7 at the 0.125 mg/ml dilutions. The immunoreactivity of caspase 6 was stronger than caspase 8 and 9. Moreover, density of caspase 6 staining was much better especially in PE 7 at the 0.125 mg/ml dilution. In conclusion, the mechanisms of apoptosis induction by PE may appear via caspase pathway because of its anticanserogenic effect. PE may be usefull in the cancer treatment protocol

Anatomy of stem cells

Human embryonic stem cells are important for the understanding of biologic mechanisms and their potential for treatment, andfor toxicology studies. Mesenchymal stem cells can produce a good environment with chemical factors and signals to serve asanti-apoptotic, immune modulator and angiogenic, and can support regeneration in damaged tissues and induce faster and better wound healing. Stem cells have recently been shown in cancer tissue; these play a major role in the progression of cancerand the principal obstacle in treatment due to their resistance. Stem cells may participate in tissue homeostasis and regenerationwhich shows their capacity for cell therapies which makes them an amazing source for the clinical translation to treatment ofdiseases. Therefore, better understanding of the behavior of these cells and the mechanisms they use are important

Cytotoxic Effect of Hypoxic Environment in Mesenchymal Stem Cell

Bone marrow mesenchymal stem cells (BM-MSCs) are used to repair hypoxic or ischemic tissue. After hypoxic the level of ATP is decreases, cellular functions do not continue and apoptosis or necrosis occur. Apoptosis is a progress of programmed cell death that occurs in normal or pathological conditions. In this study, we were investigated the hypoxic effect on apoptosis in mesenchymal stem cell. Bone marrow-derived stem cells were cultured in hypoxic (1% or 3%) or normoxic conditions 24, 96 well plates for 36 h. Cell viability was shown by MTT assay on 36 h. After fixation of cells with 4% paraformaldehyde, distributions of caspase-3, Bcl-2 and Bax with indirect immunoperoxidase technique, apoptotic cells with TUNEL assay were investigated. All staining results were evaluated using H-score analyses method with ANOVA, statistically. As a result, hypoxic condition was toxic for human mesenchymal stem cells and the number of death cell was higher in that than normoxic condition

Is Acteosid Effects on Colon Cancer Stem Cells via Inflamation and/or Apoptosis?

Colorectal cancer, as one of the main health problems worldwide. Cancer stem cells (CSCs) are called tumor-initiating cells involved in tumour heterogeneity and dormancy. CSCs may cause drug resistance, metastasis and relapse of primary and metastatic cancers. Dysregulation of survive and their cross-talk with other cell pathways may alternative way for effective treatment. We aimed whether acteosid, affects stemness and inflamatuvary prosess in primary (HCT-116) and metastatic (Colo-741) colon CSCs. CSCs were obtained from both type of colon cancer cell lines using MINIMACS system using anti-CD133. CD133+ and CD133- cells from Colo-741 and HCT-116 were cultured without or with Acteosid for 48 h. Distribution of Caspase-3, Bcl, Bax, IL-1β, TNF-α, IL-6, IL8 and IL-10 analyzed using indirect immunohistoperoxidase technique. Acteosid increased the intensity of Bax/Bcl ratio on CD133+, CD133- Colo-741 cells, there wasn’t differences of Caspase-3 immunoreactivity. Oct-4 was also decreased in CD133+ and CD133- Colo-741 cells. Caspase 3 was slightly increased CD133+ cells after incubation with Acteoside. In conclusion, it has been observed that the viability and stemness of the HCT-116 was increased after Acteoside, these properties were not changed in CD133+ Colo-741 cells. Therefore inhibition of ROS may play a role during CSCs stemness and viability

Apoptotic Properties of Rutheinum Complexes on Different Type of Cancer Cell Lines

Among chemotherapeutic agents, cisplatin and the other platinum-based drugs have occupied for 35 years an enviable position. The limitations of platinum-based drugs, dose dependent side effects and development of drug resistance mechanisms, have boosted the research for finding other metal-based drugs. Among metals, ruthenium is probably the one showing the greatest promises. Ruthenium (Ru) appears to be less toxic than platinum and several biological studies have indicated that ruthenium complexes possess diverse modes of action. The redox chemistry of ruthenium is rich and compatible with biological media, and the overall toxicity of ruthenium is lower than platinum, thus allowing higher doses of treatment. In this study we aimed that, analyses of different type of ruthenium complexes in cancer cell lines. Six Ru complexes were determined by elemental analysis, FTIR, NMR, UV-visible spectroscopy, electron density on the metal was measured by cyclic voltammetry. After that, the cellular properties of this complexes were analyses on PC-3, HT-29, Du-145 and Vero cell lines. DNA damage was analyzed H2AX staining, apoptotic cell analyses were performed flow cytometry and western blotting. After 48 h incubation of Ru complexes three of them more effective for cell lines. Especially Ru3 was more effective in cancer cell lines. Apoptotic pathway was triggered after Ru complexes incubation in PC-3, Du-145 and Ht-29 cancer cell lines. Our study suggest that Ru complexes may be used for cancer cell cytotoxicity as a drugs in patients