INCREASING THE SENSITIVITY OF CANCER CELLS KOLON WiDr 5-fluorouracil BY ETHYL ACETATE FRACTION wooden cup (Caesalpinia sappan L.) THROUGH INDUCED APOPTOSIS
ABSTRACT: Continuous administration of chemotherapy in cancer cells could decrease its sensitivity to chemotherapeutic agent. Strategy to overcome this problem is cochemotherapy with chemoprevention agent which has synergist effect with chemotherapeutic agent to eradicate cancer cells. On this research, we studied ethyl acetate fraction of Caesalpinia sappan L. (EFC) as co-chemotherapeutic agent. EFC was characterized using thin layer chromatography (TLC). Cytotoxic effect of EFC was examined using MTT assay, apoptosis detection using double staining assay, and we observed expression of cleaved PARP as apoptotic protein marker using immunocytochemistry. Potency of EFC and 5-Fluorouracil (5-FU) against WiDr cell lines was identified through IC50 value. Effect of combination 5-FU and EFC was determined form combination index (CI) as parameter of synergist effect from this combination. Apoptosis induction was determined through result from double staining assay and expression of cleaved PARP. Visual TLC profile of EFC with system consist of n-heksan, kloroform, metanol, asam asetat glasial (1:7:2:1) as mobile phase and silika F254 as stationary phase showed EFC contain brazilein as major compound with hRf 81. EFC could inhibit WiDr cancel cells growth through decrease of cells viability and morphology changing. IC50 value of EFC is 11 μg/mL in dose dependent manner. 5-FU as chemotherapeutic agent for colon cancer showed cytotoxic effect with IC50 558 μM. Combination of EFC and 5-FU showed synergist effect with CI value 0,3 at dose ½ IC50 of EFC and ½ IC50 of 5-FU. Synergist effect of this combination is predicted through apoptosis induction that showed in double staining assay result. Apoptosis mechanism was showed through decrease expression of cleaved PARP.
