ISOLASI DAN IDENTIFIKASI SENYAWA BIOAKTIF DARI KULIT BATANG Bruguiera gymnorrhiza Lamk YANG BERSIFAT SITOTOKSIK DENGAN MEKANISME APOPTOSIS PADA SEL KANKER HeLa

Cancer is one of the worldwide health issue since the incident and mortality caused by cancer continues to increase every year, one of those is cervical cancer. Treatment of cancer with chemotherapeutic drugs have not given satisfactory results and is often associated with adverse side effects and caused resistance of cancer cells, therefore searching of new anticancer compounds having new template structure from natural products continue to be carried out. Bruguiera gymnorrhiza (mangrove) is a typical coastal plant and has been used traditionally to cure gastroenteritis disease and cancer. The objectives of this study are to fractionate the methanol extract to obtain active isolate and to investigate the cytotoxic, antiproliferative and apoptosis induction activities, as well as to evaluate the expression of several proteins and to identify the structure of active isolate. Fractionation of the methanol extract of the B. gymnorrhiza stem bark was done by successive trituration using n-hexane, chloroform and ethyl acetate. Fractions were then tested their cytotoxic activities against HeLa cells. The active fraction (Chloroform fraction) was further separated by vacuum liquid chromatography using silica gel 60 H gradiently eluted with different mixture of solvents (n-hexane, chloroform, ethylacetate dan methanol). The fractions obtained were analyzed by thin layer chromatography (TLC). Fractions with similar TLC profiles are combined and tested their cytotoxic activity. Separation of the active fraction F2 gave the active fraction S3 which was further separated to give active fraction T4. Separation of T4 gave active fraction I2, which was further purified by column chromatography on sephadex LH 20 eluted with methanol yielded purple active isolate. The active isolate was tested its cytotoxic and antiproliferative activities using MTT assay, while its apoptosis induction was analyzed using double staining and flowcytometri methods, and the expression of p53, Bcl2 and caspase3 were investigated using immunocytochemistry method. The structure of active isolate was identified using UV-Vis, IR, LC-MS and NMR spectroscopic methods The result showed that chloroform fraction had cytotoxic activity with the IC50 of 134 μg/ml. The active fractions F2, S3, T4, I2, and active isolate had cytotoxic effect against HeLa cell with the IC50 of 69.4 μg/m