Elucidation of apoptotic pathway on human breast adenocarcinoma cell lines mediated by labisia pumila var. alata extract / Muhammad Faiz Zulkifli

Labisia pumila, or locally known as Kacip Fatimah were widely used by women in Malaysia to treat post-partum illnesses. Due to the increase uses of complementary medicine in breast cancer patient, this herbaceous shrub has been exploited as a replacement to the conventional therapies. Many studies i...

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Main Author:	Zulkifli, Muhammad Faiz
Format:	Thesis
Language:	English
Published:	2019
Online Access:	https://ir.uitm.edu.my/id/eprint/89716/1/89716.pdf
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spelling	my-uitm-ir.897162024-04-22T07:19:13Z Elucidation of apoptotic pathway on human breast adenocarcinoma cell lines mediated by labisia pumila var. alata extract / Muhammad Faiz Zulkifli 2019 Zulkifli, Muhammad Faiz Labisia pumila, or locally known as Kacip Fatimah were widely used by women in Malaysia to treat post-partum illnesses. Due to the increase uses of complementary medicine in breast cancer patient, this herbaceous shrub has been exploited as a replacement to the conventional therapies. Many studies indicated that Labisia pumila exerts a wide range of biological activities, including the anti-proliferation effect. However, details of the mechanism were still poorly understood. Therefore, this study was conducted to elucidate the mechanism of Labisia pumila var. alata (Lpva) inducing anti-proliferative effect towards breast adenocarcinoma (MCF-7) cell lines and the underlying mechanism. The treated cells were subjected to viability assay (MTT assay), apoptosis assay (Flow cytometry), and protein expression (Western blot). Moreover, Lpva was also tested for its estrogenic activity (Molecular docking and estrogen binding assay). Aqueous extracts from Lpva showed anti-proliferative activities in a dose- and time-dependent manner (p>0.005). Flow cytometry analysis showed that the anti-proliferative effect of Lpva induced through the apoptosis pathway where the early apoptotic cell population increase from 20 % in 24 hours to 50% in 72 hours. Molecular docking simulation showed that Lpva phytochemical able to bind to estrogen receptors with diadzein gives the lowest affinity at -9.1 for both estrogen receptors alpha and beta, while estrogen binding assay proved that aqueous extract of Lpva able to bind to estrogen receptor alpha and estrogen receptor beta. Furthermore, estrogen antagonist, fulvestrant (ICI 182,780), proved that Lpva aqueous extract exerts their anti-proliferative effect through the estrogen receptors of MCF-7 cells. Western blot analysis showed that Lpva extracts induced apoptosis through both intrinsic and extrinsic pathways. Lpva significantly increase the expression of caspase 8 and 9 at IC50 and IC75 concentration with caspase 9 showed the highest expression (1.47 protein ratio at IC50 and 1.57 protein ratio at IC75) (p>0.01). Western blot analysis also suggested that Lpva down-regulated the expression of Bcl-2 at IC75 (0.38 protein ratio) and up-regulated the expression of pro-apoptotic protein Bax at IC50 and IC75 (0.64 and 1.14 protein ratio) (p>0.01). These results elucidate the mechanism of Lpva anti-proliferative effect on MCF-7 cells. 2019 Thesis https://ir.uitm.edu.my/id/eprint/89716/ https://ir.uitm.edu.my/id/eprint/89716/1/89716.pdf text en public masters Universiti Teknologi MARA (UiTM) Faculty of Pharmacy Eshak, Zolkapli
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
advisor	Eshak, Zolkapli
description	Labisia pumila, or locally known as Kacip Fatimah were widely used by women in Malaysia to treat post-partum illnesses. Due to the increase uses of complementary medicine in breast cancer patient, this herbaceous shrub has been exploited as a replacement to the conventional therapies. Many studies indicated that Labisia pumila exerts a wide range of biological activities, including the anti-proliferation effect. However, details of the mechanism were still poorly understood. Therefore, this study was conducted to elucidate the mechanism of Labisia pumila var. alata (Lpva) inducing anti-proliferative effect towards breast adenocarcinoma (MCF-7) cell lines and the underlying mechanism. The treated cells were subjected to viability assay (MTT assay), apoptosis assay (Flow cytometry), and protein expression (Western blot). Moreover, Lpva was also tested for its estrogenic activity (Molecular docking and estrogen binding assay). Aqueous extracts from Lpva showed anti-proliferative activities in a dose- and time-dependent manner (p>0.005). Flow cytometry analysis showed that the anti-proliferative effect of Lpva induced through the apoptosis pathway where the early apoptotic cell population increase from 20 % in 24 hours to 50% in 72 hours. Molecular docking simulation showed that Lpva phytochemical able to bind to estrogen receptors with diadzein gives the lowest affinity at -9.1 for both estrogen receptors alpha and beta, while estrogen binding assay proved that aqueous extract of Lpva able to bind to estrogen receptor alpha and estrogen receptor beta. Furthermore, estrogen antagonist, fulvestrant (ICI 182,780), proved that Lpva aqueous extract exerts their anti-proliferative effect through the estrogen receptors of MCF-7 cells. Western blot analysis showed that Lpva extracts induced apoptosis through both intrinsic and extrinsic pathways. Lpva significantly increase the expression of caspase 8 and 9 at IC50 and IC75 concentration with caspase 9 showed the highest expression (1.47 protein ratio at IC50 and 1.57 protein ratio at IC75) (p>0.01). Western blot analysis also suggested that Lpva down-regulated the expression of Bcl-2 at IC75 (0.38 protein ratio) and up-regulated the expression of pro-apoptotic protein Bax at IC50 and IC75 (0.64 and 1.14 protein ratio) (p>0.01). These results elucidate the mechanism of Lpva anti-proliferative effect on MCF-7 cells.
format	Thesis
qualification_level	Master's degree
author	Zulkifli, Muhammad Faiz
spellingShingle	Zulkifli, Muhammad Faiz Elucidation of apoptotic pathway on human breast adenocarcinoma cell lines mediated by labisia pumila var. alata extract / Muhammad Faiz Zulkifli
author_facet	Zulkifli, Muhammad Faiz
author_sort	Zulkifli, Muhammad Faiz
title	Elucidation of apoptotic pathway on human breast adenocarcinoma cell lines mediated by labisia pumila var. alata extract / Muhammad Faiz Zulkifli
title_short	Elucidation of apoptotic pathway on human breast adenocarcinoma cell lines mediated by labisia pumila var. alata extract / Muhammad Faiz Zulkifli
title_full	Elucidation of apoptotic pathway on human breast adenocarcinoma cell lines mediated by labisia pumila var. alata extract / Muhammad Faiz Zulkifli
title_fullStr	Elucidation of apoptotic pathway on human breast adenocarcinoma cell lines mediated by labisia pumila var. alata extract / Muhammad Faiz Zulkifli
title_full_unstemmed	Elucidation of apoptotic pathway on human breast adenocarcinoma cell lines mediated by labisia pumila var. alata extract / Muhammad Faiz Zulkifli
title_sort	elucidation of apoptotic pathway on human breast adenocarcinoma cell lines mediated by labisia pumila var. alata extract / muhammad faiz zulkifli
granting_institution	Universiti Teknologi MARA (UiTM)
granting_department	Faculty of Pharmacy
publishDate	2019
url	https://ir.uitm.edu.my/id/eprint/89716/1/89716.pdf
_version_	1804889815621042176

Elucidation of apoptotic pathway on human breast adenocarcinoma cell lines mediated by labisia pumila var. alata extract / Muhammad Faiz Zulkifli

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