Induction of apoptosis and the signalling pathways involved by dillenia suffruticosa dichloromethane root extract in mcf-7 and mda-mb-231 breast cancer cells
Dillenia suffruticosa has been used traditionally to treat cancerous growth. Previous study reported that dichloromethane extract of D. suffruticosa root (DCM-DS) was the most cytotoxic towards breast cancer cells. The present study investigated the mode of cell death and the signalling pathways inv...
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2015
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my.upm.eprints.640492025-01-13T06:51:06Z http://psasir.upm.edu.my/id/eprint/64049/ Induction of apoptosis and the signalling pathways involved by dillenia suffruticosa dichloromethane root extract in mcf-7 and mda-mb-231 breast cancer cells Foo, Jhi Biau Dillenia suffruticosa has been used traditionally to treat cancerous growth. Previous study reported that dichloromethane extract of D. suffruticosa root (DCM-DS) was the most cytotoxic towards breast cancer cells. The present study investigated the mode of cell death and the signalling pathways involved in MCF-7 and MDA-MB-231 breast cancer cells treated with DCM-DS. DCM-DS was obtained by sequential solvent extraction. The cytotoxicity of DCM-DS was determined by using MTT assay. The mode of cell death was evaluated by using an inverted light microscope and AnnexinV/PI-flow cytometry analysis. Cell cycle analysis and measurement of intracellular reactive oxygen species (ROS) level were performed by using flow cytometry. The cells were co-treated with DCM-DS and antioxidants α-tocopherol or ascorbic acid to evaluate the involvement of ROS in the cytotoxicity of DCM-DS. Effect of DCM-DS on the expression of antioxidant, apoptotic, growth, survival genes and proteins were analysed by using GeXP-based multiplex system and Western blot, respectively. The compounds in DCM-DS were isolated by various chromatography techniques. The structure of the compounds was elucidated by using nuclear magnetic resonance analysis. DCM-DS was cytotoxic to the MCF-7 and MDA-MB-231 cells in a time-and dose-dependent manner. Cell cycle analysis revealed that DCM-DS induced G0/G1 and G2/M phase cell cycle arrest in MCF-7 and MDA-MB-231 cells, respectively. DCMDS induced apoptosis and oxidative stress in these two cell lines. Treatment with α- tocopherol reduced the cytotoxicity of DCM-DS at 50 µg/mL in the cells, suggesting that DCM-DS induced lipid peroxidation to destroy the cancer cells. Therefore, DCMDS can be employed as a pro-oxidant agent to treat breast cancer. The induction of apoptosis in MCF-7 and MDA-MB-231 cells by DCM-DS is possibly due to the activation of pro-apoptotic JNK1 and down-regulation of anti-apoptotic ERK1 and AKT1, which in turn down-regulates anti-apoptotic BCL-2 to increase the BAX/BCL-2 ratio to initiate the mitochondrial apoptotic pathway. The induction of cell cycle arrest in MCF-7 and MDA-MB-231 cells is possibly via p53/p21-dependent and p53- independent but p21-dependent pathway, respectively. A total of seven triterpene compounds were isolated. Betulinic acid (BA) appears to be the major and most cytotoxic compound in DCM-DS. Therefore, BA could be used as a mean for standardisation of herbal product from D. suffruticosa. In conclusion, the data suggest the potential application of DCM-DS in the treatment of breast cancer. 2015-04 Thesis NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/64049/1/IB%202015%2019.pdf Foo, Jhi Biau (2015) Induction of apoptosis and the signalling pathways involved by dillenia suffruticosa dichloromethane root extract in mcf-7 and mda-mb-231 breast cancer cells. Doctoral thesis, Universiti Putra Malaysia. |
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Dillenia suffruticosa has been used traditionally to treat cancerous growth. Previous study reported that dichloromethane extract of D. suffruticosa root (DCM-DS) was the most cytotoxic towards breast cancer cells. The present study investigated the mode of cell death and the signalling pathways involved in MCF-7 and MDA-MB-231 breast cancer cells treated with DCM-DS. DCM-DS was obtained by sequential solvent extraction. The cytotoxicity of DCM-DS was determined by using MTT assay. The mode of cell death was evaluated by using an inverted light microscope and AnnexinV/PI-flow cytometry analysis. Cell cycle analysis and measurement of intracellular reactive oxygen species (ROS) level were performed by using flow cytometry. The cells were co-treated with DCM-DS and antioxidants α-tocopherol or ascorbic acid to evaluate the involvement of ROS in the cytotoxicity of DCM-DS. Effect of DCM-DS on the expression of antioxidant, apoptotic, growth, survival genes and proteins were analysed by using GeXP-based multiplex system and Western blot, respectively. The compounds in DCM-DS were isolated by various chromatography techniques. The structure of the compounds was elucidated by using nuclear magnetic resonance analysis. DCM-DS was cytotoxic to the MCF-7 and MDA-MB-231 cells in a time-and dose-dependent manner. Cell cycle analysis revealed that DCM-DS induced G0/G1 and G2/M phase cell cycle arrest in MCF-7 and MDA-MB-231 cells, respectively. DCMDS induced apoptosis and oxidative stress in these two cell lines. Treatment with α- tocopherol reduced the cytotoxicity of DCM-DS at 50 µg/mL in the cells, suggesting that DCM-DS induced lipid peroxidation to destroy the cancer cells. Therefore, DCMDS can be employed as a pro-oxidant agent to treat breast cancer. The induction of apoptosis in MCF-7 and MDA-MB-231 cells by DCM-DS is possibly due to the activation of pro-apoptotic JNK1 and down-regulation of anti-apoptotic ERK1 and AKT1, which in turn down-regulates anti-apoptotic BCL-2 to increase the BAX/BCL-2 ratio to initiate the mitochondrial apoptotic pathway. The induction of cell cycle arrest in MCF-7 and MDA-MB-231 cells is possibly via p53/p21-dependent and p53- independent but p21-dependent pathway, respectively. A total of seven triterpene compounds were isolated. Betulinic acid (BA) appears to be the major and most cytotoxic compound in DCM-DS. Therefore, BA could be used as a mean for standardisation of herbal product from D. suffruticosa. In conclusion, the data suggest the potential application of DCM-DS in the treatment of breast cancer. |
| format |
Thesis |
| author |
Foo, Jhi Biau |
| spellingShingle |
Foo, Jhi Biau Induction of apoptosis and the signalling pathways involved by dillenia suffruticosa dichloromethane root extract in mcf-7 and mda-mb-231 breast cancer cells |
| author_facet |
Foo, Jhi Biau |
| author_sort |
Foo, Jhi Biau |
| title |
Induction of apoptosis and the signalling pathways involved by dillenia suffruticosa dichloromethane root extract in mcf-7 and mda-mb-231 breast cancer cells |
| title_short |
Induction of apoptosis and the signalling pathways involved by dillenia suffruticosa dichloromethane root extract in mcf-7 and mda-mb-231 breast cancer cells |
| title_full |
Induction of apoptosis and the signalling pathways involved by dillenia suffruticosa dichloromethane root extract in mcf-7 and mda-mb-231 breast cancer cells |
| title_fullStr |
Induction of apoptosis and the signalling pathways involved by dillenia suffruticosa dichloromethane root extract in mcf-7 and mda-mb-231 breast cancer cells |
| title_full_unstemmed |
Induction of apoptosis and the signalling pathways involved by dillenia suffruticosa dichloromethane root extract in mcf-7 and mda-mb-231 breast cancer cells |
| title_sort |
induction of apoptosis and the signalling pathways involved by dillenia suffruticosa dichloromethane root extract in mcf-7 and mda-mb-231 breast cancer cells |
| publishDate |
2015 |
| url |
http://psasir.upm.edu.my/id/eprint/64049/1/IB%202015%2019.pdf http://psasir.upm.edu.my/id/eprint/64049/ |
| _version_ |
1823093030385614848 |
| score |
13.250246 |