Zainuddin, Nik Aina Syazana Nik
(2016)
Elucidation of antiproliferative mechanism of dendrophthoe pentandra methanolic extract on breast cancer cells (MCF-7).
Masters thesis, Universiti Sains Malaysia.
Abstract
Cancer is one of the major causes of death, worldwide. The ultimate goal of cancer
treatment is to remove the malignant cells completely without harming healthy cells.
There is an urgency to find a safe and highly effective treatment for cancer disease
and it remains a big challenge nowadays. Throughout medical history, herbal plants
have been shown to be one of valuable sources in combating cancer such as
Dendrophthoe pentandra (DP). DP or Mistletoe is a semi-parasitic plant with
traditional claims for anticancer property. However, the mechanism underlying
anticancer activity is unclear and need to be explored. Therefore, DP was selected in
order to evaluate its antiproliferative activity and mode of cell death in cancer
treatment. The extraction of DP leaves were carried out using methanol (DPME) by
maceration technique. Phytochemicals compound present in DPME were screened
and quantified. Tannin is the most abundance phytochemical present in DPME. The
antiproliferative activities of DPME towards HeLa, HepG2, MCF-7, U-2 OS and
MDA-MB-231 cell lines have been examined by MTT Assay and IC50 values were
obtained. MCF-7 cells showed the most effective growth inhibition with lowest IC50
value upon treatment with DPME. The cytotoxicity activities towards normal kidney
MDCK and normal fibroblast L-929 cells were evaluated to determine the
cytoselectivity property of DPME. The nuclear staining by Hoechst 33258 displayed
the chromatin condensation, fragmented nuclei and formation of apoptotic bodies
upon treatment with DPME according to certain period of time. Flowcytometric
analysis using Annexin V/PI double staining has confirmed that DPME-treated
MCF-7 arrested cell cycle distribution at G1/S phase and induced apoptosis. The
mechanism of action was further confirmed by determination of protein involved in
apoptosis pathway; Bcl-2, Bax, p53 and cytochrome C. The results found out that the
increased of p53 was followed by an increment of pro-apoptotic, Bax and decreased
of anti-apoptotic, Bcl-2. Activation of Bax and inactivation of Bcl-2 triggered release
of cytochrome C which leads to apoptosis event. In conclusion, DPME demonstrated
antiproliferative activity in MCF-7 cells by induction of apoptosis. Therefore, DP has
a promising approach for breast cancer treatment.
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