INTRODUCTION Cancer is a disease caused by the growth of abnormal cells, causing damage to normal body tissues. Cancer is the second leading cause of death after cardiovascular disease globally. In approximately 9.6 million people died from cancer and about 1 of 6 deaths were due to cancer. Cancer is classified as a terminal disease because until now chemotherapy, radiotherapy, immunotherapy, or gentherapy has not given satisfactory results. Angiogenesis is a physiological process of forming new blood vessels that play an important role in the human growth and development, as well as in a wound healing. But in the development of malignant tumors or cancer angiogenesis has an important role by supplying blood for cancer cell through the formation of new blood vessels. Therefore inhibition of angiogenesis is a new hope for cancer prevention and treatment, one of them is black rice (Oryza sativa L. indica) wherein its bran contains cyanidin-3-glucoside (C3G) which has the highest anthocyanin compounds (95%). The purpose of this study is to determine the anticancer potential of black rice based in anti-angiogenesis activity using the chorioallantoic membrane (CAM) assay. MATERIAL AND METHOD PROJECT The materials needed in this research are embryonated chicken eggs, black rice, alcohol 70% and 96% purity, NaCl 0.9%, aluminum foil, aquadest, wrapping and filter paper. This research goes through several stages, among others: (1) Black rice extraction Black rice is roasted at 50ºC and then blended until it becomes a fine powder after that the maceration process is carried out. Maceration is carried out by soaking the simplicia powder with 70% ethanol, soaking for 3 days with solvent replacement every 24 hours. The maserate obtained was filtered with filter paper. After that, the extract was separated from the solvent by evaporation using a rotary evaporator at a temperature of 50ºC until a concentrated extract was obtained. (2) Chorioallantoic membrane (CAM) preparation CAM is an 8 day old embryonic chicken egg incubated in a laboratory incubator at 39°C. (3) Anti-angiogenesis activity assay The test subjects were embryonic chicken eggs divided into 6 groups of three replicates with successive concentrations 20 μg/mL, 25 μg/mL, 50 μg/mL, 60 μg/mL, 75 μg/mL. Eggs opened with how to cut egg shells into 2 parts starting from the shell close to the air cavity, after that CAM attached to the shell observed macroscopically. Observation macroscopically done with the aid of a magnifying glass. (4) Data analyze Parameters observed in the research is the number of new blood vessels or angiogenetic response to CAM after giving black rice extract. Evaluation of the antiangiogenetic effect macroscopic done with count the number of blood vessels new RESULT AND DISCUSSION Angiogenesis has an important role in the growth of cancer cells. Angiogenesis is a growth of new blood vessels (neovascularization) that allows cells cancer gets its nutrient supply and oxygen, so it can expand and metastasize. In this research, black rice extract was tested to control the angiogenesis growth by using a chorioallantoic membrane (CAM) or embryonic eggs with 8 days aged. The research was carried out using 6 different concentrations of black rice extract, namely 20, 25, 50, 60 and 75 µg/mL and control by 0.9% NaCl. The results show that the mean number of blood vessels decrease as the black rice extract concentration were rise. That were 27.6 ml; 21.0 ml; 3.67 ml; 3.0 ml; 2.0 ml. Macroscopic observations show that the higher the concentration of black rice extract, the greater the angiogenesis inhibitory effect. This can be seen from the density of the blood vessels that are formed. Black rice extract has also been shown to inhibit the MMP enzyme (2 and 9). MMP enzymes are enzymes that help cancer cells to metastasize and promote cell growth. The antioxidant in the extract of black rice has the potential as an anticancer. According to Kristamtini (2014), the dominant antioksidant substance in black rice is anthocyanin which is secondary metabolites substances. Black rice has a higher phenol and anthocyanin content compare with the other functional food ingredients. CONCLUSION This research indicates that the black rice extract has very good potential in habiting the grown of cancer cells.
Antiangiogenesis Effect and Activity of Black Rice (Oryza sativa L. indica) on Chorioalantoic Membrane (CAM) as Anticancer Agent to Inhibit Cancer Cells Invasion
Short Description:
Cancer is the second leading cause of death after cardiovascular disease globally. Angiogenesis is a physiological process of forming new blood vessels that play an important role in the human growth and development, as well as in a wound healing. But in the development of malignant tumors or cancer angiogenesis has an important role by supplying blood for cancer cell through the formation of new blood vessels. Therefore inhibition of angiogenesis is a new hope for cancer prevention and treatment, one of them is black rice (Oryza sativa L. indica) where in its bran contains cyanidin-3-glucoside (C3G) which has the highest anthocyanin compounds (95%). The purpose of this study is to determine the anticancer potential of black rice based in anti-angiogenesis activity using the Chorioallantoic Membrane (CAM) assay. This research goes through several stages, among others (1) black rice extraction, (2) chorioallantoic membrane (CAM) preparation, (3) anti-angiogenesis activity assay, and (4) data analyze. In this research, black rice extract was tested to control the angiogenesis growth by using a chorioallantoic membrane (CAM) or embryonic eggs with 8 days aged. The research was carried out using 6 different concentrations of black rice extract, namely 20, 25, 50, 60 and 75 µg/mL and control by 0.9% NaCl. The results show that the mean number of blood vessels decrease as the black rice extract concentration were rise. That were 27.6 ml; 21.0 ml; 3.67 ml; 3.0 ml; 2.0 ml.