Usnea filipendula induces apoptosis in human colon cancer cell lines

Lichens are complex organisms living in a symbiotic relationship with fungi and algae have recently received special interest in cancer research. The cytotoxic activities of Usnea filipendula Stirt. lichen extract was investigated on colon cancer cell lines, HCT-15 and HT-29. Sulphorhodamine B and ATP cell viability tests were used to monitor cytotoxic activity. The mode of cell death (apoptosis/necrosis) was determined using caspase-cleaved cytokeratin 18 (M30), caspase-3/7 activity and fluorescence staining techniques that included, Annexin-V, Hoechst 33342 and propidium iodide. Usnea filipendula showed dose and time-dependent antiproliferative effect in HCT-15 and HT-29 cells. The IC50 values in HCT-15 and HT-29 cells were 17.92 and 41.87 mu g/ml, respectively. The extract induced apoptosis in both cell lines especially in HCT-15 cells in which caspase-3/7 activity was increased. Usnea filipendula was cytotoxic to colon cancer HCT-15 and HT-29 cell lines by inducing early or late apoptosis as evidenced by translocation of phosphatidylserine, pyknotic nuclei and nuclear condensation. Further studies would help to understand the full potential of Usnea filipendula as a novel anticancer therapy

Cytotoxic effects of combination with novel synthesis palladium complex with autophagy inhibitors on prostate cancer cell lines

Prostat kanseri dünyada erkeklerde en sık rastlanan ikinci kanser türüdür. Kemoterapi tedavisine karşı gelişen direnç bu kansere bağlı ölümlerin ana nedenlerinin başında gelmektedir. Bu nedenle prostat kanseri tedavisine yönelik olarak yeni terapötik hedeflerin araştırılması halen araştırmacıların ilgi odağı olup, bu hedeflerin etkileşime girdikleri hücresel sinyal yolakları aydınlatılmaya çalışılmaktadır. Son yıllarda yapılan çalışmalar, kanser tedavisinde kullanılan metal bileşiklerin umut verici etkilerinden dolayı ilaç olarak kullanımının yaygınlaştığını göstermektedir. Çoğunlukla bir stres yanıtı ve hayatta kalma mekanizması olarak işleyen otofaji, kanser gelişimi ve tedavisi sürecinde önemli bir role sahiptir. Prostat kanseri üzerinde yapılan araştırmalar otofajinin genellikle bir hayatta kalma mekanizması olarak rol aldığını göstermektedir. Dolayısıyla bu tez çalışmasında, Palladyum (II) bileşiği [[Pd(bpma)(barb)]Cl.H2O] ile otofaji inhbitörleri (3-Metiladenin ve klorokin) kombinasyonunun insan prostat kanseri hücre soyları (LNCaP, PC-3) ve sağlıklı prostat hücre soyu (PNT1A) üzerine sitotoksik etkileri araştırılmıştır. Pd (II) bileşiğinin ve otofaji inhibitörlerinin hücre canlılığı üzerine olan etkileri MTT canlılık testi ile belirlenmiştir. Kombinasyon tedavisi (Pd (II) bileşiği ve otofaji inhibitörleri) için belirlenen dozların bu hücre soyları üzerine olan etkileri önce MTT testi ile belirlenmiştir. Daha sonra da bu etkiler ATP canlılık testi ile doğrulanmıştır. Kombinasyon tedavisinin sitotoksik etkilerinden sorumlu hücre ölümü (apoptozis/nekrozis/otofaji) mekanizmasının belirlenmesi amacıyla akım sitometri sistemi ve M30 antijen yöntemi kullanılmıştır. Hücrelerde apoptoz varlığı Hoechst 33342/Anneksin-V/Propidyum İyodür üçlü boyama yöntemi ile asidik veziküler organellerin varlığı ise akridin boyaması ile floresan mikroskopta görüntülenerek desteklenmiştir. Son olarak otofaji ve hücre ölümü ile ilişkili proteinlerin ekspresyon seviyeleri Lumineks uygulaması ve immunoblotlama yöntemi ile gösterilmiştir. Sonuç olarak, Pd (II) bileşiği ve otofaji inhibitörleri kombinasyonunun özellikle metastatik prostat kanseri hücre soyunda (PC-3) sitotoksik aktivitenin artışına ve apoptozise neden olduğu bulunmuştur. Pd (II) bileşiğinin otofaji inhibitörleri ile kullanımının yeni umut verici bir tedavi seçeneği olarak kullanılabileceği öngörüsüyle ileri in vivo deneylerin yapılması gerektiği sonucuna varılmıştır.Prostate cancer is the second most common cancer in men worldwide. The development of resistance to chemotherapy is one of the main causes of cancer death. Therefore, the search for new therapeutic agents for the treatment of cancer is still the focus of researchers with an aim to understand cellular signaling pathways. Studies in recent years have shown the widespread use of metal complexes as it has promising potential in cancer treatment. Autophagy has an important role in cancer development and treatment often as a stress response and survival mechanism. Studies on prostate cancer have shown that autophagy usually works as a survival mechanism. In this research, the cytotoxic effects of combination of palladium (II) complex [[Pd(bpma)(barb)]Cl.H2O] and autophagy inhibitors (3-methyladenine and chloroquine) are investigated on human prostate cancer cell lines (LNCaP, PC-3) and healthy prostate cell line (PNT1A). Effects of Pd(II) complex and autophagy inhibitors on cell viability were determined separately by MTT viability assay. For the combinatorial treatment ¬(Pd (II) complex and autophagy inhibitors), effects at indicated doses on cell lines were firstly determined by MTT viability assay. Then ATP viability assay was used to confirm MTT results. Flow cytometry and M30 antigen method was used to determine the mode of cell death (apoptosis/necrosis/autophagy) responsible for the cytotoxicity in combinatorial treatments. Apoptosis is supported by fluorescence images using triple staining method (Hoechst 33343, Annexin V, and Propidium iodide). Formation of acidic vesicular organelles was observed by fluorescence microscopy using acridine orange staining. Finally, protein expression levels associated to autophagy and cell death were determined by immunoblotting method and Luminex assay. As a conclusion, the combination of Pd (II) complex and autophagy inhibitors results to increased cytotoxic activity especially in metastatic prostate cancer cell line by inducing apoptosis. This combination may be a promising new treatment option. It was concluded that it is necessary to carry out further in vivo experiments

Synthesis, structures and anticancer potentials of platinum(II) saccharinate complexes of tertiary phosphines with phenyl and cyclohexyl groups targeting mitochondria and DNA

A series of new Pt(II) saccharinate complexes containing PR3 ligands (PPh3, PPh2Cy, PPhCy2 and PCy3) with progressive phenyl (Ph) replacement by cyclohexyl (Cy) were synthesized and structurally characterized by lR, NMR, ESI-MS and X-ray diffraction. The anticancer activity of the complexes was tested against human breast (MCF-7), lung (A549), colon (HCT116), and prostate (DU145) cancer cell lines as well as against normal bronchial epithelial (BEAS-2B) cells. Trans-configured complexes 1, 3 and 5 emerged as potential anticancer drug candidates. The mechanism of action of the potent complexes was then investigated in detail. The three complexes interacted with DNA by groove binding and with HSA via hydrophobic IIA subdomain. Furthermore, the complexes cleaved plasmid DNA efficiently. Cellular uptake studies in MCF-7 cells showed that the biologically active complexes were mainly localized in cytoplasm. The cytotoxic activity was a function of the lipophilicity and cellular accumulation of the complexes. As determined by M30, Annexin V and Caspase 3/7 activity assays, the complexes induced apoptosis in MCF-7 and HCT116 cells. Mechanistic studies showed that the potent complexes cause excessive generation of reactive oxygen species (ROS) and display a dual action, concurrently targeting both mitochondria and genomic DNA

Synergistic interactions between resveratrol and doxorubicin inhibit angiogenesis both in vitro and in vivo

Resveratrol is a polyphenolic compound which is found in many nutrients including grapes, peanuts, raspberries, and apples. Anti-proliferative, anti-angiogenic and apoptotic effects of resveratrol have been shown on various cancer cells. Doxorubicin is considered as one of the most effective anticancer agents and reveals its antitumor activity by induction of apoptosis and inhibition of angiogenesis. Our study reports for the first time the potent ability of resveratrol in combination with doxorubicin to inhibit angiogenesis in vitro and in vivo. The cytotoxic effect of resveratrol (1.56-100 mu M), doxorubicin (0.01-0.92 mu M) and their combination were analyzed in the human umbilical vein endothelial cells (HUVECs) by ATP assay. In vitro angiogenesis was evaluated using tube formation assay in HUVECs. In vivo anti-angiogenic activity was assessed in a chick chorioallantoic membrane (CAM) model using fertilized chicken eggs. All test groups were compared to thalidomide as a positive control, three concentrations of resveratrol (10-5-2.5 mu g/pellet) and a 2 mu g/pellet concentration of doxorubicin was examined. All data were evaluated statistically. Resveratrol and doxorubicin alone displayed inhibitory effects on angiogenesis and cell viability at higher doses. However, the combination of resveratrol and doxorubicin exhibited a significant dose-dependent inhibition of CAM angiogenesis in vivo as well as proliferation and tube formation in HUVECs compared to the positive control (+/-)-thalidomide. Our results suggest that resveratrol in combination with doxorubicin is a novel strategy in the prevention and treatment of angiogenesis