Potential chemoprevention of primary liver cancer using optimized formulation of Strobilanthus crispus (L.) blume and Centella asiatica L. juices

This study was conducted to evaluate the chemoprevention potential of Strobilanthes crispus and Centella asiatica juices on liver cancer cell line (HepG2). The first and second parts of the study were on the optimization of juices formulation and processing condition for the development of high qual...

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Main Author:	Hussin, Faridah
Format:	Thesis
Language:	English
Published:	2011
Subjects:	Liver - Cancer Cancer - Chemoprevention Vegetable juices - Research
Online Access:	http://psasir.upm.edu.my/id/eprint/31435/1/IB%202011%2021R.pdf
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spelling	my-upm-ir.314352014-05-21T01:52:24Z Potential chemoprevention of primary liver cancer using optimized formulation of Strobilanthus crispus (L.) blume and Centella asiatica L. juices 2011-08 Hussin, Faridah This study was conducted to evaluate the chemoprevention potential of Strobilanthes crispus and Centella asiatica juices on liver cancer cell line (HepG2). The first and second parts of the study were on the optimization of juices formulation and processing condition for the development of high quality juices using response surface methodology. The combined effect of two independent variables for formulation optimization i.e. herbal puree (14-24%) and honey (14-40%), and two independent variables for processing condition i.e. temperature (60-800 C) and time (8-30 min) were studied. Response surface plots showed increase in herbal and honey concentration, temperature and time of processing reduced the response variables significantly (p < 0.05). The optimum product formulation for Centella asiatica and Strobilanthes crispus juices singly were 14% of both herb and honey concentration. The optimum parameter for processing of Centella asiatica juice was at 70°C for 8 min whereas the optimum parameter for processing of Strobilanthes crispus juice was at 69°C for 8 min. The third part of the study focused on the cytotoxic activities of juices at different concentration (0.001, 0.01, 0.1, 1 and 10%) and incubation time (24, 48, 72 hours). MTT (3-(4, 5-dimethylthiazole-2yl)-2, 5-diphenyl tetrazolium bromide) assay was used to examine the cytotoxic effect of juices on Chang cell line as control and HepG2 cells. Increasing doses of both juices up to 10% had no cytotoxic effect against Chang cell line as indicated by absence of IC50 values when the cell lines were treated for up to 72 hours incubation time. However, Strobilanthes crispus juice and Centella asiatica juice inhibited the proliferation of HepG2 cancer cell lines in a concentration-dependent manner as well as exposure time. The cytotoxicity started at a concentration as low as 0.1% for both juices. At 72 hours incubation time, Strobilanthes crispus and Centella asiatica juice showed cytotoxicity effects with an IC50 of approximately 0.4 (4 mg/L) and 0.1% (1 mg/L) respectively. The fourth part of this study was to discuss on the apoptotic morphology of HepG2 cancer cells treated with both juices by quantitatively measuring the apoptotic cell percentage using flow cytometry and comet assay. Both juices induced apoptosis in a dosedependent manner after being exposed to HepG2 cell lines for 72hr. At more than 0.1% concentration of juices, both Centella asiatica and Strobilanthes crispus showed higher percentage of apoptotic cell death (sub-GI population) than the control HepG2 cells. Comet assay also showed that all cells exhibited a dose-dependent increase in DNA damage compared to controls. A significant induction of DNA damage was seen at concentrations of above 0.1% for both juices. Finally the last part of the study was to observe the changes in the c-myc, c-fos and cerbB2 oncogenes expression induced by the Centella asiatica and Strobilanthes crispus juices. Results revealed that Centella asiatica juice reduced level of c-myc but increased level of c-fos and c-erbB2 expression on HepG2 cells. Contrary, Strobilanthes crispus juices showed increased level of c-myc with reduced level of c-fos and c-erbB2 expression on HepG2 cells. In conclusion, Centella asiatica and Strobilanthes crispus juices have potential as chemopreventive agents in liver cancer cell. Liver - Cancer Cancer - Chemoprevention Vegetable juices - Research 2011-08 Thesis http://psasir.upm.edu.my/id/eprint/31435/ http://psasir.upm.edu.my/id/eprint/31435/1/IB%202011%2021R.pdf application/pdf en public phd doctoral Universiti Putra Malaysia Liver - Cancer Cancer - Chemoprevention Vegetable juices - Research Institute of Bioscience
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
topic	Liver - Cancer Cancer - Chemoprevention Vegetable juices - Research
spellingShingle	Liver - Cancer Cancer - Chemoprevention Vegetable juices - Research Hussin, Faridah Potential chemoprevention of primary liver cancer using optimized formulation of Strobilanthus crispus (L.) blume and Centella asiatica L. juices
description	This study was conducted to evaluate the chemoprevention potential of Strobilanthes crispus and Centella asiatica juices on liver cancer cell line (HepG2). The first and second parts of the study were on the optimization of juices formulation and processing condition for the development of high quality juices using response surface methodology. The combined effect of two independent variables for formulation optimization i.e. herbal puree (14-24%) and honey (14-40%), and two independent variables for processing condition i.e. temperature (60-800 C) and time (8-30 min) were studied. Response surface plots showed increase in herbal and honey concentration, temperature and time of processing reduced the response variables significantly (p < 0.05). The optimum product formulation for Centella asiatica and Strobilanthes crispus juices singly were 14% of both herb and honey concentration. The optimum parameter for processing of Centella asiatica juice was at 70°C for 8 min whereas the optimum parameter for processing of Strobilanthes crispus juice was at 69°C for 8 min. The third part of the study focused on the cytotoxic activities of juices at different concentration (0.001, 0.01, 0.1, 1 and 10%) and incubation time (24, 48, 72 hours). MTT (3-(4, 5-dimethylthiazole-2yl)-2, 5-diphenyl tetrazolium bromide) assay was used to examine the cytotoxic effect of juices on Chang cell line as control and HepG2 cells. Increasing doses of both juices up to 10% had no cytotoxic effect against Chang cell line as indicated by absence of IC50 values when the cell lines were treated for up to 72 hours incubation time. However, Strobilanthes crispus juice and Centella asiatica juice inhibited the proliferation of HepG2 cancer cell lines in a concentration-dependent manner as well as exposure time. The cytotoxicity started at a concentration as low as 0.1% for both juices. At 72 hours incubation time, Strobilanthes crispus and Centella asiatica juice showed cytotoxicity effects with an IC50 of approximately 0.4 (4 mg/L) and 0.1% (1 mg/L) respectively. The fourth part of this study was to discuss on the apoptotic morphology of HepG2 cancer cells treated with both juices by quantitatively measuring the apoptotic cell percentage using flow cytometry and comet assay. Both juices induced apoptosis in a dosedependent manner after being exposed to HepG2 cell lines for 72hr. At more than 0.1% concentration of juices, both Centella asiatica and Strobilanthes crispus showed higher percentage of apoptotic cell death (sub-GI population) than the control HepG2 cells. Comet assay also showed that all cells exhibited a dose-dependent increase in DNA damage compared to controls. A significant induction of DNA damage was seen at concentrations of above 0.1% for both juices. Finally the last part of the study was to observe the changes in the c-myc, c-fos and cerbB2 oncogenes expression induced by the Centella asiatica and Strobilanthes crispus juices. Results revealed that Centella asiatica juice reduced level of c-myc but increased level of c-fos and c-erbB2 expression on HepG2 cells. Contrary, Strobilanthes crispus juices showed increased level of c-myc with reduced level of c-fos and c-erbB2 expression on HepG2 cells. In conclusion, Centella asiatica and Strobilanthes crispus juices have potential as chemopreventive agents in liver cancer cell.
format	Thesis
qualification_name	Doctor of Philosophy (PhD.)
qualification_level	Doctorate
author	Hussin, Faridah
author_facet	Hussin, Faridah
author_sort	Hussin, Faridah
title	Potential chemoprevention of primary liver cancer using optimized formulation of Strobilanthus crispus (L.) blume and Centella asiatica L. juices
title_short	Potential chemoprevention of primary liver cancer using optimized formulation of Strobilanthus crispus (L.) blume and Centella asiatica L. juices
title_full	Potential chemoprevention of primary liver cancer using optimized formulation of Strobilanthus crispus (L.) blume and Centella asiatica L. juices
title_fullStr	Potential chemoprevention of primary liver cancer using optimized formulation of Strobilanthus crispus (L.) blume and Centella asiatica L. juices
title_full_unstemmed	Potential chemoprevention of primary liver cancer using optimized formulation of Strobilanthus crispus (L.) blume and Centella asiatica L. juices
title_sort	potential chemoprevention of primary liver cancer using optimized formulation of strobilanthus crispus (l.) blume and centella asiatica l. juices
granting_institution	Universiti Putra Malaysia
granting_department	Institute of Bioscience
publishDate	2011
url	http://psasir.upm.edu.my/id/eprint/31435/1/IB%202011%2021R.pdf
_version_	1747811613496311808

Potential chemoprevention of primary liver cancer using optimized formulation of Strobilanthus crispus (L.) blume and Centella asiatica L. juices

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