Cell viability and morphological effect of heterotrigona itama sp. (stingless bee) honey on malignant brain tumour cell line
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Date
2016
Authors
Seerangan, Priatharsine
Journal Title
Journal ISSN
Volume Title
Publisher
Pusat Pengajian Sains Perubatan, Universiti Sains Malaysia
Abstract
Introduction: Glioblastoma Multiforme (GBM), a type of primary brain tumor is
found to account for almost 80% of these tumors and, therefore, signifying the frequent
occurrence. In Malaysia, a constantly increasing incidence along with limitations on
currently available therapy options had urged the need for newer approaches to be
implemented. For centuries, honey, a sweet natural product has been described in
traditional medicine for its various medicinal uses. However, until recently honeys have
been both researched scientifically and exploited medicinally on its various kinds of
photochemical constituents.
Objectives: The study is aimed to evaluate the anticancer activity of honey sample
produced by a stingless bee species (Heterotrigona itama sp) for morphological analysis
and cytotoxic activity against malignant brain tumor cells to provide evidence on its
anticancer effect.
Methods: Human primary glioblastoma cell line; U87 and dBTRG, are grown
exponentially and treated with honey concentrations of 2-10% for three sequential days
(24h, 48h, 72h). Honey sample from the stingless bee Heterotrigona Itama sp. were
collected under sterile condition. The cytotoxicity of honey is quantified using colorimetric
MTS assay with treatment of 2-10% of stingless bee honey. The plates were incubated and
absorbance value was measured using ELISA reader at 490nm. For morphological analysis,
the cell lines were seeded into six-well plates, and treated with 2% to 10% concentration ofthe honey for 24, 48 and 72 hours. At the indicated time points, morphological changes
were examined and recorded under light microscope. Apoptosis was determined at similar
intervals and concentrations after staining the cells with a nucleic acid stain known as
Acridine Orane/Propidum Iodide.
Results: Overall, crude honey sample have higher cytotoxic activities on U87 than
dBTRG cell lines in a dose dependent manner, with U87 cell line showing higher
sensitivity towards the honey extract. For morphological analysis, honey treated cells
exhibited morphological alteration such as nuclear shrinkage, chromatin condensation and
fragmented nucleus indicating apoptotic cellular changes.
Conclusion: In conclusion, Heterotrigona Itama sp. contains an in vitro cytotoxic
activity against human glioma cell lines, therefore suggesting the anticancer effect of the
stingless bee honey. Further study is however required which includes isolation and
characterization of the anticancer agents.
Description
Keywords
Apitherapy