Ethyl acetate fraction of Garcina epunctata induces apoptosis in human promyelocytic cells (HL-60) through the ROS generation and G0/G1 cell cycle arrest: a bioassay-guided approach.
Environ Toxicol Pharmacol
; 36(3): 865-74, 2013 Nov.
Article
en En
| MEDLINE
| ID: mdl-23981377
Number of deaths due to cancer diseases is increasing in the world. There is an urgent need to develop alternative therapeutic measures against the disease. Our study reports the cytotoxicity activity of Garcina epunctata (gutifferae) in human promyelocytic leukemia cells (HL-60) and prostate cancer cells (PC-3) was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Changes in mitochondrial membrane potential (MMP), reactive oxygen species (ROS) and morphological changes associated with apoptosis were examined by flow cytometry and Hoescht staining respectively. The results of in vitro antiproliferative screening of fractions and extract from G. epunctata indicated that three fractions inhibited the viability of PC-3 cells with IC50 varied from 50 to 88 µ/ml while two fractions inhibited the proliferation of HL-60 cells with IC50 range between 47.5 and 12 µg/ml. Among the entire fraction tested, Hex-EtOAc (75:25) showed cytotoxic effects on the two cell lines and EtOAc fraction was most active only HL-60 cells (12 µg/ml). Treatment of HL-60 cells with G. epunctata (20, 50, 100 µg/ml) for 24 h led to a significant dose-dependent increase in the percentage of cells in sub-G1 phase by analysis of the content of DNA in cells, and a number of apoptotic bodies containing nuclear fragments were observed in cells treated with 100 µg/ml. The EtOAc fraction of G. epunctata treatment significantly arrested HL-60 cells at the G0/G1 phase (p<0.05) and ROS was significantly elevated as well as the loss of membrane mitochondrial potential in a concentration dependant manner. The results demonstrated that the EtOAc fraction of G. epunctata inhibited the proliferation of HL-60 cells, leading to cell cycle arrest and programmed cell death, which was confirmed to occur through the mitochondrial pathway.
Palabras clave
2',7'-dichlorodihydrofluorescin diacetate; 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide; Apoptosis; Cytotoxicity; DCFH(2)-DA; G. epunctata; HL-60 cells; Hex; Hex-EtOAc; MMP; MTT; Mitochondrial membrane potential; PI; ROS; Rh-123; Rhodamine-123; hexane; hexane etyl acetate; mitochondrial mitochondrial potential; propidium iodide
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Extractos Vegetales
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Ciclo Celular
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Fase de Descanso del Ciclo Celular
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Fase G1
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Especies Reactivas de Oxígeno
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Apoptosis
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Garcinia
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Acetatos
Límite:
Humans
Idioma:
En
Revista:
Environ Toxicol Pharmacol
Año:
2013
Tipo del documento:
Article
Pais de publicación:
Países Bajos