RESUMO
OBJECTIVE: Plants belonging to the genus Celastrus exhibit antitumor activity and the ability to reverse multidrug resistance in tumor cells; however, it remains unclear whether the compound oleanen from Celastrus hypoleucus also exhibits antitumor activity. The objective of this study was to explore the inhibitory effect of 12-oleanene-3ß, 6α-diol (oleanen) on the proliferation of cervical cancer HeLa cells in vitro, as well as its relative mechanism. METHODS: HeLa cells were treated with different concentrations of oleanen for different times. Cell proliferation was determined by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide assay. Cell apoptosis was evaluated by flow cytometry and caspases activities assay. The expression of several proapoptotic proteins belonging to the Bcl-2 family, such as Bax, Bim, and Bad, was detected by Western blot. RESULTS: Oleanen mainly inhibited the proliferation of HeLa cells at the G0 to G1 and G2 to M phases, and the IC50 of oleanen for cells was significantly higher at 24 hours compared to 48 hours (17.45 ± 3.71 vs 9.02 ± 0.83 µg/mL, respectively; P < 0.05). The significant increase in activity of caspase 3/7, caspase 6 in oleanen-treated HeLa cells indicated that oleanen promoted the apoptosis of HeLa cells. The activity of caspase 9 representing the endogenous apoptotic pathways also increased obviously in oleanen treatment. Furthermore, the increase in the expression of Bim was the most significant among the Bcl-2 family after oleanen treatment. CONCLUSION: Oleanen up-regulates the expression of Bim and other proapoptotic molecules to activate the endogenous apoptosis pathway, thus promoting apoptosis and inhibiting proliferation of human cervical cancer HeLa cells in vitro.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Proteínas Reguladoras de Apoptose/metabolismo , Apoptose/efeitos dos fármacos , Celastrus , Proteínas de Membrana/metabolismo , Ácido Oleanólico/análogos & derivados , Extratos Vegetais/farmacologia , Proteínas Proto-Oncogênicas/metabolismo , Neoplasias do Colo do Útero/metabolismo , Antineoplásicos Fitogênicos/isolamento & purificação , Proteína 11 Semelhante a Bcl-2 , Western Blotting , Caspases/metabolismo , Proliferação de Células/efeitos dos fármacos , Feminino , Citometria de Fluxo , Células HeLa , Humanos , Concentração Inibidora 50 , Ácido Oleanólico/isolamento & purificação , Ácido Oleanólico/farmacologia , Extratos Vegetais/isolamento & purificação , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Regulação para Cima , Neoplasias do Colo do Útero/patologiaRESUMO
AIM: The aim of this study was to investigate whether RNA interference (RNAi) targeting vascular endothelial growth factor (VEGF) could inhibit the proliferation and induce apoptosis of ovarian carcinoma cells. METHODS: Human epithelial ovarian carcinoma cell line CaoV3 was transfected with VEGF-targeted small interfering RNA (siRNA) for 48 h. The down-regulation of VEGF expression was determined by real-time polymerase chain reaction and Western blot. The proliferation of CaoV3 cells was detected by a colorimetric BrdU assay. Caspase-3 activity and TUNEL assay were also detected to study the apoptosis of ovarian carcinoma cells induced by VEGF siRNA. The protein levels of survivin, MMP2 and MMP9 were measured by Western blot. RESULTS: mRNA and protein of VEGF were significantly down-regulated by VEGF siRNA. Down-regulation of VEGF expression dramatically suppressed the proliferation of CaoV3 cells, and increased the Caspase-3 activity. Nearly 100% of cells indicated TUNEL-positive. The expression of anti-apoptotic protein survivin was decreased, and the invasive related protein MMP2 and MMP9 were also significantly reduced by VEGF siRNA. CONCLUSION: Our study implied that siRNA targeting VEGF could effectively inhibit cell proliferation, induce cell apoptosis, and decrease the cell invasive potential. These findings suggest that the RNAi approach targeting VEGF may be an effective therapeutic strategy for ovarian cancer.
Assuntos
Apoptose , Proliferação de Células , RNA Interferente Pequeno/farmacologia , Fator A de Crescimento do Endotélio Vascular/genética , Análise de Variância , Western Blotting , Linhagem Celular Tumoral , Células Cultivadas , Regulação para Baixo , Feminino , Humanos , Marcação In Situ das Extremidades Cortadas , Proteínas Inibidoras de Apoptose , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Ovário/citologia , Interferência de RNA , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Survivina , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
OBJECTIVE: To evaluate the effectiveness of microwave endometrial ablation (MEA) in the treatment of menorrhagia in patients with severe systemic disease or medical conditions. METHODS: Forty-two menorrhagic women undergoing systemic disorders with failure of medical management were treated with MEA under local or general anesthesia, and were followed-up for 1 year. RESULTS: The women had a mean age of 39.4 years (range, 17-49). The procedure was successfully completed in all patients, and no intraoperative complications occurred. Two cases died of their primary severe medical diseases within 2 months of treatment but these cases were not associated with MEA. Among the remaining 40 patients, 24 (60.0%) had amenorrhea within 12 months. The duration of hospitalization and the amount of blood transfusion were significantly reduced after treatment, and the quality of life of these patients was improved significantly. CONCLUSIONS: MEA is a safe and effective treatment for the management of severe menorrhagia in patients undergoing systemic illness or severe medical conditions.