Cell Death Effects Of Garcinia Atroviridis Extracts On Dbtrg-05mg And U-87mg Human Brain Cancer Cell Lines
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Date
2017-08
Authors
Hashim, Bebe Syakinah
Journal Title
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Publisher
Universiti Sains Malaysia
Abstract
The aim of this study was to investigate the cell death effects of the essential oils extracted from the leaf (EO-L) and stem bark (EO-B) of G. atroviridis on DBTRG-05MG and U-87 MG human brain cancer cells. The anti-proliferative effects of EO-L and EO-B were determined using MTT assay by microplate reader. The mode of cell death induced by EO-L on U-87 MG cell line was determined using Annexin-V FITC and propidium iodide staining assay by flow cytometry. Whilst autophagy activity was determined using NVP-BEZ235 inhibitor assay by microplate reader. The effect of EO-L on mitochondrial membrane potential of U-87 MG cells was determined using JC-1 staining assay by fluorescence microscopy and fluorescence microplate reader.
Based on the MTT assay result, EO-L potentially inhibited both DBTRG-05MG and U-87 MG cells in comparison to EO-B. The growth inhibition induced by EO-L was significantly observed in U-87 MG when compared to DBTRG-05MG cells. The cell death induced by EO-L on U-87 MG cells was via apoptosis. At 24 h post-treatment, apoptosis was significantly induced in U-87 MG cells treated with EO-L at all concentration tested in comparison to the untreated cells. The loss of mitochondrial membrane potential is an early indicator for the mechanism of apoptotic cell death. EO-L has induced the loss of mitochondrial membrane potential in U-87 MG cells. The findings herein showed that U-87 MG cells were stained with JC-1 green fluorescence, which indicated the loss of mitochondrial membrane potential, and this observation was correlated with the readings obtained by fluorescence microplate reader. This study also found that co-treatment of EO-L and NVP- BEZ235 inhibitor potently induced the autophagic cell death in U87 cells.
In conclusion, EO-L potently inhibited the growth of U-87 MG human brain cancer cells, by inducing the loss of mitochondrial membrane potential which ultimately leads to the apoptotic and autophagic cell death
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Keywords
Cell death effects of the essential oils , extracted from the leaf and stem bark.