title: 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.
abstract: 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.
authors: Department of Physiological Sciences and Biochemistry, Faculty of Medicine and Biomedical Sciences, University of Yaoundé I, PO Box 1364 Yaoundé, Cameroon; Cancer Pharmacology Division, Indian Institute of Integrative Medicine, 180001, Canal Road, Jammu, India. Electronic address: apieme@yahoo.fr.
authors: Pieme
authors: P
authors: Constant Anatole
authors: Santoh Kumar
authors: SK
authors: Guru
authors: Ngamegni
authors: N
authors: Bathelemy
authors: Ngogang
authors: N
authors: Jeanne
authors: Shashi
authors: S
authors: Bhushan
authors: Tetsuya
authors: T
authors: Murayama
authors: Ajit Kumar
authors: AK
authors: Saxena
mesh_terms: D000085:Acetates / Q000737:chemistry*; D017209:Apoptosis / Q000187:drug effects*; D001681:Biological Assay; D006690:Bisbenzimidazole; D002453:Cell Cycle / Q000187:drug effects*; D004396:Coloring Agents; D004273:DNA, Neoplasm / Q000096:biosynthesis; D016193:G1 Phase / Q000187:drug effects*; D029760:Garcinia / Q000737:chemistry*; D018922:HL-60 Cells; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D024301:Plant Bark / Q000737:chemistry; D010936:Plant Extracts / Q000494:pharmacology*; D018547:Plant Stems / Q000737:chemistry; D017382:Reactive Oxygen Species / Q000378:metabolism*; D016192:Resting Phase, Cell Cycle / Q000187:drug effects*; D012997:Solvents
chemical_list: D000085:Acetates; D004396:Coloring Agents; D004273:DNA, Neoplasm; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D012997:Solvents; C007650:ethyl acetate; D006690:Bisbenzimidazole
keywords: 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