Induction of Early Apoptosis and Reactive Oxygen Species (ROS) Production by Tanzanian Basidiomycete ( Cantharellus Miomboensis )
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Date
2010
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Abstract
Cantharellus miomboensis is a new basidiomycete fungus recently found in Miombo woodlands in
Tanzania. In this study, crude extract was prepared from fruiting bodies of C. miomboensis and was in vitro
screened for its cytotoxicity using Tetrazolium salt (3-(4,5-dimethlthiazol-2-yl)-2,5-diphenyl tetrazolium
bromide (MTT) in human cell lines namely; Hepatocellular carcinoma (HepG2), Human non-small cell lung
carcinoma (H157) and Human colon adenocarcinoma (HT.29). Thereafter, pro-apoptotic effects were
determined using biochemical changes in apoptotic cells. These included externalization of phospholipid
phosphatidylserine (PS) using APO Percentage dye by flow cytometry and depolarization of mitochondrial
membrane potential using Tetramethyl rhodamine ethyl ester perchlorate (TMRE) assay. The test extract was
found to induce dose dependent PS externalization on human cell lines when treated with various
concentrations (1 - 5 mg/ml) and completely depolarized the mitochondrial membrane potential after 6 hours
on HepG2 cell line. When the extract was examined for ROS production using 2’,7’- dichlorofluorescin
diacetate (DCFH-DA) staining, there was no ROS generation found in HepG2 cells. It is therefore concluded
that C. miomboensis extract is able to induce apoptosis in HepG2 cells and PS externalization and loss of
mitochondrial membrane potential in HepG2 cells appear to be independent of ROS production.
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Keywords
Miombo woodlands, Apoptosis, Basidiomycete, Reactive oxygen species, Carcinoma
Citation
Masalu, R., Hosea, K.M., Meyer, M., Lyantagay, S. and Kanyande, S., 2010. Induction of early apoptosis and reactive oxygen species (ROS) production by Tanzanian basidiomycete (Cantharellus miomboensis). International Journal of Biological and Chemical Sciences, 4(4).