| Summary: | Cervical cancer is placed at the fourth most frequent cancer among women
worldwide, which was estimated approximately 530, 000 new cases in 2012.
This type of cancer has become one of the leading causes of cancer death among
women worldwide. Conventional and modern cancer treatment nowadays comes
with negative and adverse side effects to the patients. Therefore, lot of studies
have been conducted to search for new alternative treatment which is more safe
and effective such as utilisation of natural product based medications. Previous
studied revealed various pharmacological activities of Quercus Infectoria (QI)
galls or manjakani including anticancer activity. However, the mechanism of
action lay behind was not well explained. Therefore, in this present study, QI
gall was selected for the evaluation of cytotoxic activity and cell death mode of
action. In this study, QI was extracted using different types of organic solvents
before being tested for cytotoxicity activity and mode of cell death. MTT assay
was used to determine cytotoxic activity for n-hexane (QJH), ethyl acetate
(QIEA) and methanol (QIM) of QI galls extracts against cervical cancer (Hela).
Cytotoxic activity of QI extracts also tested against normal fibroblast (L929)
cell line to determine cytotoxic effects and cytoselective properties. Meanwhile,
DMSO-treated cells served as negative control while cisplatin-treated cells
served as positive control. After that, dose-response graph was plotted to
determine IC50 values. Observation on nuclear morphology, apoptotic percentage and detection of apoptotic protein expressions were also conducted.
Moreover, phytochemical screening analysis had been carried out to determine
bioactive groups that present in the most potent extract. From the findings, QIEA extract exhibited better cytotoxic activity with best growth inhibition
against Hela cells (IC50 value= 6.33 ± 0.33 µg/ml) compared to other extracts
and showed cytoselective property as no cytoxicity observed in the treated
normal fibroblast (L929) cells. As for the phytochemical analysis of QIEA
extract, it was revealed the presence of tannin, alkaloids, glycosides, saponins,
terpenoids, flavonoids and phenolic compounds The QIEA extract was then
purified and resulted two compounds 4-0 methylgallic acid (MGA) and gallic
acid (GA). The most potent antiproliferative activity was exhibited by MGA
sub fraction with IC50 II ±0.58 Jlg/ml. The current study also revealed that Hela
cells treated with MGA subfraction has undergone apoptosis as exerted by the
alteration of nuclear morphology and the presence of apoptotic bodies as well as
the increment of apoptosis rate in the treated cells. Furthermore, MGA
subfraction triggered apoptosis through upregulation of tumor suppression
protein p53 , with down regulation of Bcl-2 expression and facilitated the
apoptosis execution through caspase-3 activation . In conclusion, the purification
of QIEA has resulted two biologicaly active subfraction namely MGA and GA.
It was cleared that MGA subfraction reduced Hela cell growth through
induction of apoptosis.
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