Activities Kuasinoid semisynthetic compound of Fruit Makasar (Brucea javanica [L.] Merr) as Anticancer with Target Protein P53, Bcl-2, caspase-3, COX-2 and c-Myc
ABSTRACT: The Incidence of cervical cancer recently had a leading caused of cancer in woman mostly in developing country including Indonesia. Therefore the pursuit of that cancer drugs is still needed. Trial in vitro models of cervical cancer by using HeLa cell has been approved. It has been known the role of several gens in cancer development which include P53, Bcl-2, c-Myc and Caspase-3. Due to apoptosis DNA repaired was influenced by P53 proteins complex. Cancer development also depends on inflammatory effect contributed by COX-2. Brucea javanica (L.) Merr as a herbal plant has been used to treat several diseases as antiinflamatory, antibabesial, antivirus, antiplasmodial as well as anticancer. In this study it was performed how to isolate bruceine A from the seed of “buah makasar”. Following had semi-synthesis of that compound with each of chlorobenzoyl chloride, and benzoyl chloride. As a result bruceine A have been isolated (0.0035%), 3-O-chlorobenzoylbruceine (38%) and 3-O-benzoyl bruceine (40%). The cytotoxic activity and its toxicity were done both in HeLa and Vero cells. That isolated and semisynthesis products confirmed by structural elucidation with UV/vis, FT-IR, H-NMR, CNMR, HPLC, HRESI-MS and LC-MS. Molecular activity was tested by flow cytometry and immunocytochemistry assay through induction apoptosis by increasing P53 and caspase-3 levels, antiproliferation through the increasing of P53 expression and the lowering levels among of Bcl-2, c-Myc, as well as COX-2. The IC50 value of bruceine A on HeLa cells as cytotoxicity effect were 87.45±13.0 μg/mL; 3-O-chlorobenzoylbruceine 41.5±4.79 μg/mL and 3-O-benzoylbruceine 119.12±2.65 μg/mL with doxorubicin 5.66±1.22 μg/mL and vinblastin 8.77±2.32 μg/mL as positive control. The IC50 value of bruceine A, 3-O-chlorobenzoylbruceine, 3-O-benzoylbruceine, doxorubicin and vinblastin on Vero cells tested were more selective than HeLa cells. Their cytotoxicity effect were 1366.55±53.43 μg/mL; 1385.15±81.57 μg/mL; 870.61±4.00 μg/mL; 61.46±28.37 μg/mL; and 102.43±16.86 μg/mL respectively. That isolated and semisynthesis compounds gave in 100, 50, 25 ìg/mL serial concentration have increased the P53 expression (93.93%-45.13%), decreased Bcl-2 expression (81.19%-18.99%); increased Caspase-3 (cleaved) (96.81%-31.06%) and decreased the c-Myc expression (23.08%-70.93%); decreased COX-2 expression (19,60%- 9,01%). This study comes to conclusions that bruceine A, 3-O-chlorobenzoylbruceine, and 3-Obenzoylbruceine showed anticancer cervix activity via apoptotic inductions, inhibiting of their proliferation and inflammatory effects.
