Expression of caspase-8 BREAST CANCER CELLS INDUCED EXTRACT OF T47D lime leaves (Citrus hystrix DC.)
ABSTRACT: Breast cancer is the second most common cancer among women in Indonesia. This cancer mostly treated with chemotherapy. Chemotherapy treatment not only cause side effect to patient but also induce resistance to the cancer cells. Kaffir lime (Citrus hystrix DC.) contains bioactive compounds which act as anti-cancer. Our previous study showed that kaffir lime leaves had cytotoxic effect and induced apoptosis in T47D. Apoptosis is regulated by caspase cascade such as caspase-8, initiator of caspase. Objective of this study was to determine the expression of caspase-8 in T47D cells after treated with ethyl acetate and chloroform extract of kaffir lime leaves. The methods include leaves extraction of kaffir lime leaves with maceration method, treatment on T47D cells, RNA isolation of T47D cells, cDNA synthesis, house keeping gene (GAPDH) and caspase-8 amplification. Cells were treated with extract for 24 hours. The concentration of the extract based on IC50 of each extract, which were 283 μg/ml and 129 μg/ml for ethyl acetate and chloroform, respectively. Whereas concentration of doxorubicin as positive control was 5μg/ml. The results showed that the cells were given doxorubicin, ethyl acetate and chloroform extract expressed the caspase-8 with the bands at 211bp. The band of caspase-8 which were expressed by doxorubicin sample was thinner than ethyl acetate and chloroform extract samples. The presence of caspase-8 in the extracts sample showed that apoptosis in T47D cells have been activated. Hence, extract lime leaves can be used as an alternative medicine for cancer treatment.
