RESUMO
OBJECTIVE: The objective of this study was to determine the cytotoxic effect and apoptotic activity of Rhaponticum carthamoides transformed root (TR) and root of soil-grown plant (NR) extracts in a human glioma primary cells. The effect of these root extracts on cell cycle arrest, mitochondrial membrane potential (ΔΨm) and expression levels of apoptosis-related genes (Bcl-2, Bax and p53) were also examined. METHODS: Cytotoxic activity of root extracts was evaluated by MTT assay. Apoptosis and cell cycle were determined by flow cytometry. Expression levels of apoptosis-related gene were analysed by RT-PCR and Western blotting. ΔΨm was examined by the use of JC-1 reagent. KEY FINDINGS: Rhaponticum carthamoides root extracts inhibit cell growth and induce apoptosis in a dose-dependent manner in human glioma cells. The root extracts were found to up-regulate the pro-apoptotic Bax protein and down-regulate the anti-apoptotic Bcl-2 protein, consequently increasing the ratios of Bax/Bcl-2 protein levels. Moreover, an increase of the p53 protein level and reduction of ΔΨm in glioma cells were observed after treatment with NR and TR extracts. CONCLUSION: The results of this study may offer a new insight into the potential anticancer activity of R. carthamoides root extracts.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias Encefálicas/tratamento farmacológico , Proliferação de Células/efeitos dos fármacos , Glioma/tratamento farmacológico , Leuzea/química , Extratos Vegetais/farmacologia , Raízes de Plantas/química , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Proteína X Associada a bcl-2/metabolismo , Agrobacterium/fisiologia , Antineoplásicos Fitogênicos/isolamento & purificação , Apoptose/efeitos dos fármacos , Western Blotting , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Glioma/genética , Glioma/metabolismo , Glioma/patologia , Leuzea/microbiologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Microscopia de Fluorescência , Fitoterapia , Extratos Vegetais/isolamento & purificação , Raízes de Plantas/microbiologia , Plantas Medicinais , Proteínas Proto-Oncogênicas c-bcl-2/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais/efeitos dos fármacos , Transformação Bacteriana , Proteína Supressora de Tumor p53/genética , Proteína X Associada a bcl-2/genéticaRESUMO
Russian knapweed (Rhaponticum repens) is a rangeland weed pest in the western United States. One candidate fungus for biological control of R. repens is Puccinia acroptili, which causes a rust disease. Understanding the life cycle of candidate rust fungi for weed biological control is an essential component in risk assessments and evaluations, and for P. acroptili such was unknown. For this reason greenhouse studies were undertaken to clarify the life cycle of P. acroptili under artificial conditions. Spermogonia with spermatia developed on R. repens following plant inoculation with teliospores. Artificial transfer of spermatia between spermogonia resulted in the development of aecia with uredinioid aeciospores. Inoculation with aeciospores or urediniospores resulted in uredinia containing urediniospores and occasional amphispores. Telia with teliospores and occasional mesospores developed later. Teliospores produced typical basidia with four basidiospores. These results suggest that the life cycle of P. acroptili is macrocyclic and autoecious. Inoculation with teliospores also frequently resulted in production of sori that were morphologically similar to aecia but which were not associated with spermogonia or the classical transfer of spermatia. The ontology of these sori is unknown. This is the first description of spermogonia and the first report and description of basidiospores, aecia, aeciospores, amphispores and mesospores of P. acroptili.
