CYTOTOXIC ACTIVITY OF CRUDE AQUEOUS-METHANOL EXTRACT AND FRACTIONS OF Ulva fasciata AND ITS EFFECTS ON RELEASE OF CYTOCHROME C FROM MITOCHONDRIA IN HUMAN BREAST CANCER CELL LINE (MCF-7)

• Main Author: DWI SUSANTI (NIM : 20515036), MAULIDDINA
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/23008
• Institution: Institut Teknologi Bandung

Cancer is a group of diseases involving abnormal cell growth with the potential to invade to other parts of the body. Breast cancer is the second leading cause of cancer death in women worldwide. Cancer cells are resistant to apoptosis and show a shift in energy production from mitochondrial oxidative phosphorylation to cytosolic glycolysis. Mitochondria have two essential role that is for cell survival by energy production and to initiate apoptosis by changes in the permeability of mitochondrial membrane and the release of pro apoptotic molecules such as cytochrome c into the cytoplasm. Cytochrome c was released from mitochondria leading to the onset of apoptosis. Bioactive components have been suggested to induce apoptosis. More natural products contain bioactive components that have selective anti-cancer activity, that only kills cancer cells but not normal cells. Based on FAO data, Indonesia is the third largest country as a producer of seaweed after China and Philippines. Ulva fasciata is a common green seaweed has been known the potential for traditional cancer drugs. A previous study showed that a crude aqueous-methanol extract and active fractions of Ulva fasciata could induce apoptosis in isolated mouse liver cells characterized by cytochrome c release from mitochondria at 5.68 &#956;M or equivalent to 15.1% of 500 &#956;M mitoxantrone and 10.06 &#956;M or equivalent to 26.7% of 500 &#956;M mitoxantrone. Isolation of an active fraction showed the presence of phenolic, glycosides and terpenoids compounds. Extract of Ulva fasciata also induced apoptosis in colon cancer cells (HCT116). <br /> <br /> <br /> The aim of the present study was to investigate the cytotoxic activities of a crude aqueous-methanol extract and active fractions of Ulva fasciata against MCF-7 cancer cells, so that later can be known its ability as anti-cancer agent. This study also investigate effects of crude extract and active fractions of Ulva fasciata treatment againts release of cytochrome c from mitochondria of MCF-7 as one of the symptoms of apoptosis. Cytotoxic activity was evaluated by MTT assay and calculation of IC50 value and concentration of cytochrome c released from mitochondria was calculated after treatment. <br /> <br /> <br /> The results showed that Ulva fasciata contained 78.78% (w/w wet sample) water. <br /> <br /> <br /> An extraction process of active compound from Ulva fasciata using the maceration in methanol solvent p.a: chloroform p.a: 50 mM phosphate buffer with pH 7.6. (2: <br /> <br /> <br /> 1: 0.5 v/v). The percentage extract yield was for crude aqueous-methanol extract (ECMU) is 0.96% (w/w dry sample). Crude extracts separated into 10 major fractions by column chromatography and grouped based on similar UV-Vis spectra were collected from 200 to 400 nm, which include wavelengths of phenolic and flavonoids compounds. F4, F6 and F9, respectively, had the greatest TPC (288.19; 144.89; 263.89 mg GAE/g dried samples) and TFC (152.78; 107.95; 142.16 mg QE/g dried sample) and they were tested for their antitumor activity against MCF-7 cancer cells. The result obtained with MTT assay showed that IC50 of F6 (24.33 &#956;g/ml) and F9 (20.52 &#956;g/ml) met the American National Cancer Institute standard, were more cytotoxic than EKMU (33.94 &#956;g/ml) and F4 (31.37 &#956;g/ml. MCF-7 cells were treated with 1 &#956;g/ml doxorubicin released cytochrome c with concentration of 21.465 &#956;M. MCF-7 cells treated with EKMU, F6 and F9 in IC50 could release cytochrome c, respectively, adalah 1,376 &#956;M or equivalent to 6,41% doxorubicin 1 &#956;g/ml ; 2.327 &#956;M or equivalent to 10,84%doxorubicin 1 &#956;g/ml ; 8.127 &#956;M or equivalent to 37.86% doxorubicin 1 &#956;g/ml. It can be concluded that F6 and F9 Ulva fasciata had cytotoxic compounds and induce to release of cytochrome c from mitochondria in MCF-7 cells. The result of a spectrophotometric wavelength analysis showed that F9 had absorptionmaximum at 207, 211 and 273 nm, while F6 had absorption maximum at 207, 224, 261, 286 and 325 nm, which include absorbance of phenolic and flavonoids compounds.