Biomaterial and Biomedicine, 2013, Vol.2, No.1, 1-5
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3
atmosphere with 5% CO
2
.
Cytotoxity of the crude
extract II, III, A, B, C and D were measured using
MTT {(3-(4, 5-dimethylthiazol-2-yl)-2, 5-dipheny-
ltetrazolium bromide (Sigma, St. Louis, USA)} assay.
For this, the cells were grown at a concentration of 5×
10
3
cells/well in 96 well plates.
After 24 h, cells were washed with fresh medium
and were treated with different concentrations of
crude extract (12.5 µg/mL, 25 µg/mL, 50 µg/mL and
100
µg/mL). The cells without the addition of crude
extract were taken as control. After 24 h incubation,
100
µL of MTT (1 mg/mL) solution was added and
further incubated for 4 h at 37 C. Finally 100 µL
DMSO was added to solubilize the formazan salt
formed and the amount of formazan salt was
determined by measuring the OD at 540 nm using a
GENios ® microplate reader (Tecan Austria GmbH,
Austria). The relative cell viability was determined by
the amount of MTT converted into formazan salt.
Viability of cells was quantified as a percentage
compared to that of control. The experiment was
carried out in triplicate and the data were expressed as
mean from these three sets of experiments.
3.
Results and Discussion
In the present study, the ability of the compounds in
the sixteenth fraction of the extracts of the sponge
Aurora globostellata
to inhibit the proliferation of
human HeLa, Raw 264.7 and HEK-293 cell line was
examined. Results from cell counting showed that the
compound, at all the doses tested, inhibited the
proliferation of cells. In addition, the HeLa, Raw
264.7
and HEK-293 cell line viability was estimated
by MTT assay, and the results showed that the
compounds in
Aurora globostellata.
These results were expressed as percent viability and
as total number of viable cells As shown in Figure 1,
the extract was cytotoxic at concentrations of 12.5
g/mL and 100 g/mL. Viability decreased following a
dose/time response curve. The anticancer property of
cell free extracts from sponge isolates might be due to
the presence of the active secondary metabolites such
as alkaloids and quninine (Gordaliza, 2010). Alkaloids
are microtubule interfering agents which can bind with
beta tublin, thus preventing the cell from making the
Figure 1 Anti cancer activity of
Aurora globostellata
marine
sponges
mitotic spindle fibres necessary to move the
chromosome around as the cell divides(Solanki et al.,
2008),
inhibiting topoisomerase (Facompre et al.,