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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 2013; 36(3):865-74ET

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+Show Affiliations

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.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23981377

Citation

Constant Anatole, Pieme, et al. "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." Environmental Toxicology and Pharmacology, vol. 36, no. 3, 2013, pp. 865-74.
Constant Anatole P, Guru SK, Bathelemy N, et al. 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. 2013;36(3):865-74.
Constant Anatole, P., Guru, S. K., Bathelemy, N., Jeanne, N., Bhushan, S., Murayama, T., & Saxena, A. K. (2013). 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. Environmental Toxicology and Pharmacology, 36(3), pp. 865-74. doi:10.1016/j.etap.2013.07.015.
Constant Anatole P, et al. 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. 2013;36(3):865-74. PubMed PMID: 23981377.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Constant Anatole,Pieme, AU - Guru,Santoh Kumar, AU - Bathelemy,Ngamegni, AU - Jeanne,Ngogang, AU - Bhushan,Shashi, AU - Murayama,Tetsuya, AU - Saxena,Ajit Kumar, Y1 - 2013/08/06/ PY - 2013/04/22/received PY - 2013/07/18/revised PY - 2013/07/26/accepted PY - 2013/8/29/entrez PY - 2013/8/29/pubmed PY - 2014/7/16/medline KW - 2′,7′-dichlorodihydrofluorescin diacetate KW - 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide KW - Apoptosis KW - Cytotoxicity KW - DCFH(2)-DA KW - G. epunctata KW - HL-60 cells KW - Hex KW - Hex-EtOAc KW - MMP KW - MTT KW - Mitochondrial membrane potential KW - PI KW - ROS KW - Rh-123 KW - Rhodamine-123 KW - hexane KW - hexane etyl acetate KW - mitochondrial mitochondrial potential KW - propidium iodide SP - 865 EP - 74 JF - Environmental toxicology and pharmacology JO - Environ. Toxicol. Pharmacol. VL - 36 IS - 3 N2 - 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. SN - 1872-7077 UR - https://www.unboundmedicine.com/medline/citation/23981377/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_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1382-6689(13)00162-2 DB - PRIME DP - Unbound Medicine ER -