RESUMO
In the current study we investigated the inhibitory effect of rucaparib (Rubraca®) on human ovarian cancer SKOV3 and A2780 cells and its possible mechanism. Cancer cells and human normal ovarian epithelial IOSE80 cells were treated with Rubraca® at different concentrations. Cell viability was measured by MTT assay. Necrotizing apoptosis was detected by Annexin V-FITC/PI double staining combined with flow cytometry. Reactive oxygen species were measured by 2',7'-dichlorofluorescent yellow diacetate (DCFH-DA) fluorescent probe. The expression of receptor-interacting protein kinase 1 (RIP1) and RIP3 protein was determined by Western Blot. Our data showed that Rubraca® inhibited the proliferation of ovarian cancer SKOV3 and A2780 cells in a dose-and time-dependent manner. After Rubraca® treatment, the apoptotic rate of SKOV3 and A2780 cells (Annexin V+/PI-cells) did not change significantly, but the proportion of necrotic cells (PI+cells or Annexin V+/PI+cells) increased significantly, which was different from the control group. Furthermore, Rubraca® could significantly induce SKOV3 and A2780 cells to produce excessive reactive oxygen species and significantly upregulate the expression of RIP1 and RIP3. When pretreated with reactive oxygen species inhibitor N-acetyl-L-cysteine (NAC) or RIP1 inhibitor (Nec-1), the necrosis apoptotic rate of SKOV3 and A2780 cells decreased significantly. In summary, Rubraca® could significantly inhibit the proliferation of ovarian cancer SKOV3 and A2780 cells, which may be partially achieved via upregulating the expression of RIP1 and RIP3 proteins, and activating the process of necrotic apoptosis.
Assuntos
Apoptose/efeitos dos fármacos , Indóis/farmacologia , Neoplasias Ovarianas/tratamento farmacológico , Inibidores de Poli(ADP-Ribose) Polimerases/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Humanos , Indóis/administração & dosagem , Necrose/tratamento farmacológico , Complexo de Proteínas Formadoras de Poros Nucleares/genética , Neoplasias Ovarianas/genética , Inibidores de Poli(ADP-Ribose) Polimerases/administração & dosagem , Proteínas de Ligação a RNA/genética , Espécies Reativas de Oxigênio/metabolismo , Proteína Serina-Treonina Quinases de Interação com Receptores/genética , Fatores de Tempo , Regulação para Cima/efeitos dos fármacosRESUMO
The seasonal dynamics of the total flavonoid contents in various parts of Dryopteris erythrosora, a traditional Chinese medicinal fern, and their antioxidant activity were investigated. The total flavonoids content in various parts of D. erythrosora showed an obvious seasonal dynamic change. The total flavonoid contents in stems (from 4.3% to 12.5%) were much higher than that in leaves with an average content of 2.01%. In spring, the total flavonoid contents in stems were relatively low, but increased rapidly from summer to winter. However, the seasonal dynamics of total flavonoid contents in leaves showed different model. The total flavonoid contents in the stems showed a negative correlation with that in the leaves from January to July. The correlation coefficient of about -0.7 was obtained. The antioxidant activity of the extracts also altered in proportion to the change of total flavonoid contents. In general, the extracts from stems always showed highest antioxidant potentials and it was suggested that the stems can be used as crude medicine.