APOPTOSIS INDUCED BY MICROMINUTININ ON HUMAN BREAST CANCER CELLS, MCF7
M.A. Abdah, R. Asmah, Y.Y.H Taufik , Patimah I & Fauziah O
Abstract
APOPTOSIS INDUCED BY MICROMINUTININ ON HUMAN BREAST CANCER CELLS, MCF7.
M.A. Abdah1*, R. Asmah2, Y.Y.H Taufik3 , Patimah I1 & Fauziah O1
1Department of Biomedical Sciences, 2Department of Nutrition and Health Sciences, Faculty of Medicine and Health Sciences, 3Department of Chemistry, Faculty of Science and Environmental Health, Universiti Putra Malaysia, 43400 Serdang, Selangor Malaysia.
Treatment such as surgery, chemotherapy and radiotherapy are currently being used for cancer patients. Recently, a new approach, gene therapy, is being considered. Another approach is by using the anti-cancer properties of natural products. Micromelum minutum Wight et Am (Rutaceae), is a tall shrub, common in Malaysia and widespread in South-East Asia. The plant is traditionally used in the treatment of fever and giddiness. Not many studies were done on the effect of extracts of Micromelum minutum on cancer. The purpose of this study is to analyze the effect of a new compound, microminutinin isolated from Micromelum minutum on MCF7, human breast cancer cell line. The cell line was maintained in tissue culture flask at 37°C in an atmosphere of 5% CO2 in complete RPMI 1640 media. A stock solution was prepared and a range of concentration was treated into MCF7. Microculture tetrazolium assay (MTT assay) was utilized to assess the cytotoxic effect of these concentrations. IC50 of microminutinin was obtained from the cell viability graph. Based on the cell viability graph, the ICSO of microminutinin against the human breast cancer cell line, MCF7 was 203.0 ± 4.0 IM. The mode of cell death by microminutinin was studied using the acridine orange and propidium iodide staining. MCF7 cells treated with the compound were left for 48 hours and were then stained with 10 ig / ml acridine orange and 10 ig / ml propidium iodide. Cells without treatment served as negative control. The morphological changes were viewed under confocal microscope. After 48 hours, cell shrinkage with nucleus condensation was observed. For confirmation, a test using Annexin V/propodium iodide was used. The compound-treated cell lines were maintained for 24 and 48 hours. They were then fixed and treated with Annexin V/propodium iodide and scanned using the flow cytometer. The results showed that after 24 and 48 hours, 0.2 ± 0.14% and 3.6 ± 0.28% of apoptotic cells were detected, respectively. This study verified that microminutinin from Micromelum minutum induces apoptosis in hormone dependent breast cancer cell line, MCF7. However, more studies on the mechanism of the anticancer action is recommended.
*Corresponding authour: Abdah@medic.upm.edu.my Tel : +6(03)89468447
