Protein Profile Breast Cancer Cells T47D After Giving Kaffir Lime Leaf Extract (Citrus hystrix DC.)
ABSTRACT: Kaffir lime (Citrus hystrix DC.) leaves contains flavonoid, coumarin, saponin and terpenoid. Previous study showed that kaffir lime leaves are cytotoxic and can induce apoptosis of cancer cells. Giving chemoprevention compound cause differences in certain protein expression. Dose and exposure time affect the effectiveness of chemoprevention agents. The objective of this study was to examine the effect of ethyl acetate and chloroform extracts of kaffir lime leaves with a time dependent to protein profile of T47D (human breast cancer cell line) and Vero. Vero are noncancerous cells were used for comparison in chemoprevention agent test. The methods include leaves extraction, protein cells extraction, measuring the concentration by Bradford assay and separation of protein bands by SDS-PAGE. Cells treated with different exposure time 12, 24 and 48 hours. Dose of the extract is based on the IC50 value of each cell. Doxorubicin at a dose of 2.5 mikrog / ml and 5 mikrog / ml was used as a positive control. The molecular weight of protein samples was determined from linear regression equation of log protein marker molecule weight to the value of Rf (retention factor). Results showed that the value of T47D cell protein molecular weight range from 111.4-10.4 kDa and Vero cell range from 9,3-111,2 kDa. The time dependent cause different protein profiles in T47D and Vero cells, as well as treatment with extract of ethyl acetate and chloroform. Analysis of the protein profile by SDS-PAGE can be used as a preliminary study to test the effect of lime leaves more specific as cancer drugs.
